Monoclonal antibody that specifically binds to GITR

Abstract

The present invention relates to the field of biotechnology, and particularly to antibodies or their antigen-binding fragments, and to the use thereof. More particularly, the present invention relates to monoclonal antibodies that bind specifically to GITR. The invention also relates to a nucleic acid that codes for said antibody or for an antigen-binding fragment thereof, an expression vector, a method for producing the antibody, and the use of said antibody for treating diseases or disorders associated with GITR.

Description

antibodies or to use More specifically, the present antibodies that specifically bind to GITR TNFR-related protein / INFRSF18 / tumor factor superfamily, member 18). The invention also relates to a encoding said antibody or antigen-binding fragment expression vector, a method for preparing said antibody, and use said antibody in treatment of diseases or disorders associated GITR. Background of the invention TNFRSF18, GITR (Glucocorticoid-induced protein / INFRSF18 / tumor necrosis factor receptor superfamily) membrane protein, a receptor belonging to tumor necrosis receptor superfamily. GITR is a type - transmembrane protein consisting of 216 acids and having a molecular weight of 26 kDa. The extracellular domain comprises three TNFR-Cys repeats and glycosylation site. The three cysteine-rich domains and tail of GITR share significant homology with 4-1BB, (Nocentini, et al. (1997) Proc. Natl. Acad. Sci. 94:6216-6221). Human GITR is expressed at low levels in responder T cells, CD4+ cells exhibiting increased expression relative to CD8+ GITR expression 1s significantly up-regulated for several following T cell activation. GITR is constitutively expressed at levels in regulatory T cells (Tregs), such as CD4+CD25+ or cells, and is further up-regulated when these cells are (Nocentini and Riccardi (2005) E. J. Immunol. 35:1016). However, GITR expression is not exclusively limited to T Several works have also indicated that GITR is expressed on NK macrophages, B cells, dendritic cells, mast cells and (Nocentini and Riccardi (2005) E. J. Immunol. 35:1016-1022). GITR is expressed in lymph nodes, peripheral blood white cells and to a lesser extent in the spleen, constitutively in high amounts on Tregs, in low amounts on naive T cells cells. However, GITR is expressed not only on immune cells, but tumor cells. RNA-Seq data analysis in relation to GITR 33 tumor types has been conducted and has revealed that expressed in HNSCC (head and neck squamous cell carcinoma), small cell lung cancer), breast cancer, esophageal cancer cancer. RNA-Seq analysis of samples from 24 tumor types has shown results. Thus, GITR is expressed not only on immune cells, on the membrane of tumor cells. In tumor samples, GITRL-Fc the gene expression associated with T cells, CD8 T cells, Thl cells, interferon gamma, NK cells, Teff cells, T activation markers. Expression of GITR and ligand thereof is not haematopoietic cells. GITR 1s also expressed on osteoclast precursors, whereas GITRL 1s expressed on cells. GITRL is a type II transmembrane protein as is typical for TNF ligand family members. Current research indicates that human typically exists as a trimer, although it can also be present as monomer or assemble into other multimeric forms (Chattopadhyay, et (2007) Proc. Natl. Acad. Sci. 104:19452-19457; Zhou, et al. Proc. Natl. Acad. Sci. 105:635-640). There is some evidence that a soluble form of GITRL is also produced (Baltz, et Blood 112:3735-3743; Mahesh, et al. (2006) Eur. J. Immunol. 36: 2138). GITRL is expressed primarily on antigen presenting cells including macrophages, B cells, dendritic cells and endothelial that can function as APC (Nocentini and Riccardi (2005) E. J. 35:1016-1022; Agostini, et al. (2005) Infect. Immun. 73:7502-7508; Nocentini, et al. (2007) E. J. Immunol. 37:11651169). Binding of GITRL on APC to GITR on responder T cells GITR signaling, which co-stimulates responder T suppressive activity of regulatory T cells. GITR as a co-activating signal to both CD4+ and CD8+ inducing or enhancing proliferation and particularly when T cell receptor (TCR) stimulation is close to (Schaer, et al. (2012) Curr. Opin. Immunol. 24:217224). specifically, GITR can have several effects on effector T cells regulatory T cells, including: co-stimulation and effector T cells such that they are more resistant to inhibition of regulatory T cells, decreasing of the T cells to suppression by regulatory T cells and of regulatory T cells from the circulation (Nocentini, et Eur. Immunol. 37:1165-1169). functions of GITR, and thus of are to enhance proliferation T cells, inhibit the suppressive effect of are devoid of the ability to tumor and suppression by Treg cells. GITR at secondary stages of priming and expansion, through anti-GITR antibody, soluble GITR ligand or DC vaccine modulates Teff and Treg tumor responses in favor of the former, thus tumor regression. Thus, anti-GITR antibody provides Teff to Treg suppression. Collectively, the foregoing functions, in particular costimualtion of responder T cells and abrogation of the activity of regulatory T cells, means that GITR activation an enhanced immune response. Such activation has the restore immune responses to infections and to tumors. molecules capable of activating GITR would be of immunostimulatory agents under conditions when it is desirable trigger an enhanced immune response. The antibody must possess the properties of a GITR agonist, effector, cytotoxic properties against Treg lymphocytes. Various antibodies against GITR are known in the W02015187835, WO02015031667, wW02017068186, W02017214548) . 23 anti-GITR agonists (antibodies / recombinant GITRLs) are now preclinical and clinical trials. Only two antibodies are in Phase clinical trials (TRX518, INCAGN1876), and MEDI1873, are in Phase 1 clinical trials. Very few are However, at the moment, no antibody that GITR and that is approved for therapeutic use the In connection with the above, it is te agonistic antibodies, that interact with GITR, abrogate the suppressor effect of regulatory T cells, the T-suppressor (regulatory) component of the immune system ADCC effector properties. BCD-166 is an agonist monoclonal antibody that interacts GITR, activates the receptor, abrogates the suppressor regulatory T cells, inhibits / deletes the T-suppressor component of the immune system through ADCC effector thereby increasing the number of CD8+ and CD4+ effector activating the T-effector component of the immune system microenvironment. Summary In one aspect, the present invention relates to an monoclonal antibody or antigen-binding fragment thereof specifically binds to GITR comprising: (a) a heavy chain variable domain comprising: (1) CDR1 comprising an amino acid sequence selected from group: NYGMH (SEQ ID NO: 1) or YYWMY (SEQ ID NO: 12); (ii) CDR2 comprising an amino acid sequence selected from group: VIWFDGSNKFYTDSVKG (SEQ ID NO: 2) or AISWNGGRTYYAESMKG (SEQ 13); (iii) CDR3 comprising an amino acid sequence selected from group: ELGGYYYDSSGFRPYYYGMDV (SEQ ID NO: 3) or NRYYSDPNYGMNL (SEQ 14), and (b) a light chain variable domain comprising: (1) CDR1 comprising an amino acid sequence selected from group: RASQSIGSWLA (SEQ ID NO: 7) or TGTSTDIGTYKYIS (SEQ ID NO: 17); (ii) CDR2 comprising an amino acid sequence selected from group: AASTLOR (SEQ ID NO: 8) or GVSHRPS (SEQ ID NO: 18); (iii) CDR3 comprising an amino acid sequence selected from group: QQSHSHPLT (SEQ ID NO: 9) or SSYTSSGTVV (SEQ ID NO: In some embodiments, a monoclonal antibody or fragment thereof comprises a heavy chain variable CDR 1, 2 and 3 comprising amino acid sequences sequences of ID NOs: 1, 2 and 3, respectively. In some a monoclonal antibody or fragment thereof a heavy chain variable CDR 1, 2 and 3 amino acid sequences sequences of SEQ ID 13 and 14, respectively. In some embodiments, a monoclonal antibody or fragment thereof comprises a light chain variable CDR 1, 2 and 3 comprising amino acid sequences represented by NOs: 7, 8 and 9, respectively. In some embodiments, a monoclonal antibody or fragment thereof comprises a light chain variable CDR 1, 2 and 3 comprising amino acid sequences represented by NOs: 17, 18 and 19, respectively. In some embodiments, a monoclonal antibody or fragment thereof comprises: . a heavy chain variable domain comprising CDRs 1, 2 comprising amino acid sequences represented by the sequences ID NOs: 1, 2 and 3, respectively; a light chain variable domain comprising CDRs 1, 2 comprising amino acid sequences represented by the sequences ID NOs: 7, 8 and 9, respectively. In some embodiments, a monoclonal antibody or fragment thereof comprises: a heavy chain variable domain comprising CDRs 1, 2 comprising amino acid sequences represented by the sequences ID NOs: 12, 13 and 14, respectively; a light chain variable domain comprising CDRs 1, 2 comprising amino acid sequences represented by the sequences ID NOs: 17, 18 and 19, respectively. In some embodiments, a monoclonal antibody or fragment thereof comprises a heavy chain variable an amino acid sequence that is at least 90% homologous to acid sequence of SEQ ID NO: 4. In some embodiments, a monoclonal antibody or fragment thereof comprises a heavy chain variable the amino acid sequence of SEQ ID NO: 4. In some embodiments, a monoclonal antibody or fragment thereof comprises a heavy chain variable an amino acid sequence that is at least 90% homologous to acid sequence of SEQ ID NO: 15. In some embodiments, a monoclonal antibody or fragment thereof comprises a heavy chain variable the amino acid sequence of SEQ ID NO: 15. In some embodiments, a monoclonal antibody or fragment thereof comprises a light chain variable an amino acid sequence that is at least 90% homologous to acid sequence of SEQ ID NO: 10. In some embodiments, a monoclonal antibody or fragment thereof comprises a light chain variable the amino acid sequence of SEQ ID NO: 10. In some embodiments, a monoclonal antibody or fragment thereof comprises a light chain variable an amino acid sequence that is at least 90% homologous to acid sequence of SEQ ID NO: 20. In some embodiments, a monoclonal antibody or fragment thereof comprises a light chain variable the amino acid sequence of SEQ ID NO: 20. In some embodiments, a monoclonal antibody or fragment thereof comprises: - a heavy chain variable domain comprising an amino acid that is at least 90% homologous to the amino acid sequence NO: 4; - a light chain variable domain comprising an amino acid that is at least 90% homologous to the amino acid sequence of NO: 10. In some embodiments, a monoclonal antibody or fragment thereof comprises: - a heavy chain variable domain comprising the amino of SEQ ID NO: 4; - a light chain variable domain comprising the amino of SEQ ID NO: 10. In some embodiments, a monoclonal antibody or fragment thereof comprises: - a heavy chain variable domain comprising an amino acid that is at least 90% homologous to the amino acid sequence of NO: 15; - a light chain variable domain comprising an amino acid that is at least 90% homologous to the amino acid sequence of NO: 20. In some embodiments, a monoclonal antibody or fragment thereof comprises: - a heavy chain variable domain comprising the amino of SEQ ID NO: 15; - a light chain variable domain comprising the amino of SEQ ID NO: 20. In some embodiments, a monoclonal antibody that binds to GITR is a full-length IgG antibody. In some embodiments, the monoclonal IgG antibody is of human IgG2, IgG3 or IgG4 isotype. - In some embodiments, the monoclonal IgG antibody is of human isotype. In some embodiments, a monoclonal antibody that binds to GITR comprises the E345R mutation in the Fc increase agonist properties, antibody-dependent cellular (ADCC), but not complement-dependent cytotoxicity (CDC). In some embodiments, a monoclonal antibody comprises comprising an amino acid sequence that is at least 90% homologous the sequence of SEQ ID NO: 5. In some embodiments, a monoclonal antibody comprises comprising the amino acid sequence of SEQ ID NO: 5. © In some embodiments, a monoclonal antibody comprises comprising an amino acid sequence that is at least 90% homologous the sequence of SEQ ID NO: 6. In some embodiments, a monoclonal antibody comprises comprising the amino acid sequence of SEQ ID NO: 6. © In some embodiments, a monoclonal antibody comprises a light comprising an amino acid sequence that is at least 90% homologous the sequence of SEQ ID NO: 11. In some embodiments, a monoclonal antibody comprises a light comprising the amino acid sequence of SEQ ID NO: 11. In some embodiments, a monoclonal antibody comprises: - a heavy chain comprising an amino acid sequence that is at 90% homologous to the sequence of SEQ ID NO: 5; - a light chain comprising an amino acid sequence that is at 90% homologous to the sequence of SEQ ID NO: 11. In some embodiments, a monoclonal antibody comprises: - a heavy chain comprising the amino acid sequence of SEQ ID 5; - a light chain comprising the amino acid sequence of SEQ ID 11. In some embodiments, a monoclonal antibody comprises: - a heavy chain comprising an amino acid sequence that is at 90% homologous to the sequence of SEQ ID NO: 6; - a light chain comprising an amino acid sequence that is at 90% homologous to the sequence of SEQ ID NO: 11. In some embodiments, a monoclonal antibody comprises: - a heavy chain comprising the amino acid sequence of SEQ ID 6; - a light chain comprising the amino acid sequence of SEQ ID 11. In some embodiments, a monoclonal antibody comprises comprising an amino acid sequence that is at least 90% homologous the sequence of SEQ ID NO: 16. In some embodiments, a monoclonal antibody comprises comprising the amino acid sequence of SEQ ID NO: 16. © In some embodiments, a monoclonal antibody comprises a light comprising an amino acid sequence that is at least 90% homologous the sequence of SEQ ID NO: 21. In some embodiments, a monoclonal antibody comprises a light comprising the amino acid sequence of SEQ ID NO: 21. In some embodiments, a monoclonal antibody comprises: - a heavy chain comprising an amino acid sequence that is at 90% homologous to the sequence of SEQ ID NO: 16; - a light chain comprising an amino acid sequence that is at 90% homologous to the sequence of SEQ ID NO: 21. In some embodiments, a monoclonal antibody comprises: - a heavy chain comprising the amino acid sequence of SEQ ID 16; - a light chain comprising the amino acid sequence of SEQ ID 21. In one aspect, the present invention relates to an nucleic acid that encodes any said antibody or antigen-binding thereof. In some embodiments, a nucleic acid is DNA. In one aspect, the present invention relates to an vector comprising the above nucleic acid. In one aspect, the present invention relates to obtaining a host cell to produce said antibody or fragment thereof, which comprises transformation of a with vector. In one aspect, the present invention relates to a host cell preparing said antibody or antigen-binding fragment thereof said nucleic acid. In one invention relates to a method or antigen-binding fragment host cell in a culture medium said if necessary, aspect, the present invention relates to a used for treating a disease or disorder said antibody or antigen-binding fragment effective amount in combination with one or excipients. In some a pharmaceutical composition is intended be used for or disorder mediated by GITR from the group cervical cancer, head and neck stomach cancer, breast cancer, renal cell CRC cancer), (OC) ovarian cancer, NSCLC (non-small In one aspect, the present invention relates to a composition for treating a disease or disorder comprising said antibody or antigen-binding therapeutically effective amount and at least one active antitumour compound in a therapeutically In some embodiments, a pharmaceutical is be used for treating a disease or disorder GITR from the group comprising cervical cancer, head neck stomach cancer, breast cancer, renal cell cancer, CRC cancer), (OC) ovarian cancer, NSCLC (non-small cell lung cancer). In some embodiments, a pharmaceutical composition comprises therapeutically active antitumour compound selected from chemotherapeutic agent, antibody or anti-hormonal agent. In some embodiments, a pharmaceutical composition therapeutically active antitumour compound that is an selected from the group comprising: anti-PDl1 antibodies, antibodies, anti-CTLA4 antibodies, anti-4-1BB antibodies, antibodies or combinations thereof. In some embodiments, a pharmaceutical composition therapeutically active antitumour compound that is a small In some embodiments, a pharmaceutical composition therapeutically active antitumour compound selected from the group activators of innate or adaptive immunity. In some embodiments of a pharmaceutical composition, antibody and at least one therapeutically active antitumour are administered sequentially. In some embodiments of a pharmaceutical composition, antibody and at least one therapeutically active antitumour are administered simultaneously. In one aspect, the present invention relates to a method inhibiting the biological activity of GITR in a subject in need such inhibition, comprising administering an effective amount of antibody or antigen-binding fragment thereof. In one aspect, the present invention relates to a treatment of a disease or disorder mediated by GITR administering to a subject in need of such treatment said antigen-binding fragment thereof or said pharmaceutical in a therapeutically effective amount. In some embodiments, a method for treatment includes a or disorder selected from the group comprising: cervical cancer, and neck cancer, stomach cancer, breast cancer, renal cell (colorectal cancer), (OC) ovarian cancer, NSCLC (non-small cancer) . In one invention relates the use of composition for treatment in a subject in need of such treatment of disease or disorder mediated by GITR. In some embodiments, the use includes a disease or selected from the group: cervical cancer, head and neck cancer, cancer, breast cancer, renal cell cancer, CRC (colorectal (OC) ovarian cancer, NSCLC (non-small cell lung cancer). Brief description of drawings Fig. 1. Plasmid for protein production pEE-GITR-TEV-Fc lama. Fig. 2. Plasmid for protein production pEE-GITR -Fc. Fig. 3. Map of plasmid pEE-humGITR-ligand-foldon-EPEA. Fig. 4. SDS-PAGE (4-20% gel) 1. Actemra 2.5 mg 2. Fermentas unstained PW marker 3. I’ GITR-TEV-Fc-lama +B-ME 10 ml 5. Angiopoetin 2-H6F + R-ME 10 ml 6. 7. GITR-TEV-Fc-lama -f -ME 10 ml 8. Angiopoetin 2-H6F - B-ME 10 ml Fig. 5. SDS gel electrophoresis 1. Fermentas unstained PW marker 5 3. hGITR ligand -EPEA medium before purification 5 ml I’ hGITR ligand -EPEA medium after purification 5 ml 5. hGITR ligand -EPEA Fig. 6. Scheme of synthesis of human naive combinatorial Fig. 7. Map of phagemid pHS5 used for cloning Fab phage libraries. Fig. 8. Map of expression plasmid pLL for production of Fabs. Fig. 9. Map of expression vector pEE-BCD166-01-001-VH-HC the heavy chain of antibodies for transient antibody mammalian cells Fig. 10. Map of expression vector encoding the light chain of antibodies production in mammalian cells Fig. 11. SDS-PAGE in 12% GEL + B-ME 1. Actemra 5 ml 2. Fermentas unstained PW marker 3. 4. anti-GITR antibody lot#1161 medium before purification ml 5 anti-GITR antibody lot#1161 medium after purification 5 6. anti-GITR antibody lot#1161 5 ml 7. anti-GITR antibody lot#1162 5 ml 8. anti-GITR antibody lot#1163 5 ml 9. anti-GITR antibody lot#1164 5 ml 10. anti-GITR antibody lot#1165 5 ml 11. anti-GITR antibody lot#1166 medium before purification ml 12. medium after purification 5 13. 5 ml Fig. 12. SDS-PAGE in 12% GEL + B-ME 1. Fermentas unstained PW marker Zs anti-GITR antibody lot#1208 medium before purification ml 3. anti-GITR antibody lot#1208 medium after purification ml 4 anti-GITR antibody lot#1208 5 ml 5. anti-GITR antibody lot#1209 5 ml 6. anti-GITR antibody lot#1210 5 ml 7. anti-GITR antibody lot#1211 5 ml 8. anti-GITR antibody lot#1212 5 ml 9. anti-GITR antibody lot#1213 5 ml 10. anti-GITR antibody lot#1214 medium before purification ml 11. anti-GITR antibody lot#1214 medium after purification ml 12. anti-GITR antibody lot#1214 5 ml Fig. 13. A graph showing the results of of agonist activity of anti-GITR monoclonal (BCD166-01-01), 1210 BCD166-01-011) and reporter cell line assay. Fig. 14. A graph showing the results of examination of binding of BCD166-01-01 antibody to human GITR, mouse cynomolgus monkey GITR. Fig. 15. A graph showing the results of examination of binding of BCD166-01-011 antibody to human GITR, mouse cynomolgus monkey GITR. Fig. 16. A graph showing the results of examination of binding of BCD166-01-014 antibody to human GITR, mouse cynomolgus monkey GITR. Fig. 17. A graph showing the results of level of activation in cell agonist assay of BCDl66-01-01, BCD166-02-01, BCD166-01-014 question. Fig. 18. A graph showing the results of EC50 value in ADCC of BCD166-02-01, BCD166-01-01, BCD166-01-014, BCD166-02-014 question. Fig. 19. A graph showing the results of CDC level of 01, BCD166-02-01, BCD166-01-014 in question. Fig. 20. A graph showing the results of the effect of candidates in question on responder cells (NK) as compared to control. Fig. 21. A graph showing the results of effect of candidates in question on responder cells (B) as compared to 22. A graph showing the results of effect of in question on responder cells (CD3) as compared to 23. A graph showing the results of effect of in question on responder cells (CD8+ T cells) as to control. 24. A graph showing the results of effect of in question on responder cells (CD4+ T cells) as to control. nTreg depletion Fig. 26. Analysis nTreg depletion effect of anti-GITR donor 2 cell Big. 27. of depletion of iTregs effect of for donor © and donor cell The results of analysis of effect of of secretion of proinflammatory IL-2 and t+he anti-cancer effect. Table of data for binding affinity constants of 02-01 to gamma receptors. A graph showing the stability of BCD166-02-01 under hour at 50 °C. Fig. 31. A graph showing the stability of BCD166-01-014 under hour incubation at 50 °C. Fig. 32. Mean values of tumor volume in groups during experiment. Fig. 33. Index of tumor growth inhibition on day 33. Description of the invention Definitions and general methods Unless defined otherwise, all technical and scientific terms herein have the same meaning as 1s commonly understood by one ordinary skill in the art. Further, unless otherwise required by context, shall include pluralities and plural terms shall include Typically, the classification and methods of cell culture, biology, immunology, microbiology, genetics, analytical organic synthesis chemistry, medical and pharmaceutical well as hybridization and chemistry of protein and described herein are well known and widely used by those the art. Enzyme reactions and purification methods are according to the manufacturer's instructions, as is common in or as described herein. Definitions associated with the antibody TNFRSF18, GITR (Glucocorticoid-induced protein / INFRSF18 / tumor necrosis factor receptor superfamily) membrane protein, a receptor belonging to tumor necrosis receptor superfamily. GITR is a type I transmembrane protein of 216 amino acids and having a molecular weight of 26 kDa. The terminal extracellular domain comprises three TNFR-Cys repeats N-glycosylation site. The three cysteine-rich domains and tail of GITR share significant homology with 4-1BB, (Nocentini, et al. (1997) Proc. Natl. Acad. Sci. 94:6216-6221). Amplification of the GITR gene and / or overexpression of thereof were found in many cancer diseases, including: cervical head and neck cancer, stomach cancer, breast cancer, renal cell CRC (colorectal cancer), (OC) ovarian cancer, NSCLC (non-small lung "binding molecule" includes antibodies term (Ig), as used and antigen chains heavy (H) chains and two light (L) chains interconnected by bonds, or an antigen-binding portion. Each heavy chain heavy chain variable region (abbreviated referred to herein as a heavy chain constant region. Known are five types of mammalian heavy chain denoted by Greek letters: «, &, &, vy and up. The of heavy chain present defines the class of an antibody; these found in IgA, IgD, IgE, IgG, and IgM antibodies, respectively. heavy chains differ in size and composition; o and vy approximately 450 amino acids, while nu £ have amino acids. Each heavy chain has two the variable region. The constant region of the same isotype, but differs in of different Heavy chains vy, «o and & have a constant region composed of constant domains CH1, CH2 and CH3 (in a line), and a hinge region added flexibility (Woof J., Burton D., Nat Rev Immunol 4, 2004, 99); heavy chains pu and e have a constant region composed of constant domains CH1, CH2, CH3 and CH4. In mammals, known are types of light chain denoted by lambda (A) and kappa (x). Each chain consists of a light chain variable region (abbreviated to herein as VL) and light chain constant region. The length of a light chain is 211 to 217 amino acids. light chain is a kappa (x) light chain, and the constant preferably C kappa (xX). "Antibodies" to can be of any class IgA, IgD, IgE, IgG, or subclass (e.g. IgG2, IgG3, IgG4, IgAl VL and VH regions regions called interspersed between that are more conserved, regions (FR). Each VH and VL is composed of three CDR and arranged from amino-terminus to carboxy-terminus in the order: FR1, CDR1l, FR2, CDR2, FR3, CDR3, FR4. The variable the heavy and light chains contain a binding domain that with an antigen. The constant regions of the antibodies the binding of the immunoglobulin to host tissues or factors, various cells of the immune system (e.g. effector cells) and the component (Clg) of the classical complement system. The term "antigen-binding portion" of an antibody or binding fragment" (or simply "antibody portion" or fragment"), as used herein, refers to one or more fragments antibody that retain the ability to specifically bind to It has been shown that the antigen-binding function of an by fragments of a full length fragments which are included within the term of an antibody include (i) Fab-fragment of the VL, VH, CL and CH 1 domains; (ii) F(ab') 2 a fragment comprising two Fab-fragments linked by a at hinge region; (iii) Fd- consisting of the and (iv) Fv-fragment of the VL and VH of a arm of an antibody; (v) aly, Nature which consists of a extracted determining region In regions VL VH, are can be methods and VH region are paired to form monovalent molecules chain Fv (scFv); see e.g. Bird et al. (1988) Science Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). assumed that such single-stranded molecules are also included the term "antigen-binding portion" of an antibody. Such fragments are obtained using conventional techniques known to skilled in the art, and the fragments are screened in the same as are intact antibodies. Preferably, the CDR of antigen-binding portion or antibody antigen binding portion of the invention is mouse, lama or human donor library or substantially of with certain amino acid residues altered, e.g. different amino acid residues in order to optimize the specific antibodies, e.g. KD, koff, IC50, EC50, the framework regions of antibodies of the invention are of origin or substantially of human origin (at least 80, 85, 90, 95, 97, 98 or 99% of human origin). In other embodiments, the antigen binding portion of the may be derived from other non-human species including mouse, rabbit, rat or hamster, but not limited to. Alternatively, the binding region can be derived from the human species. The term "variable" refers to the fact that certain portions the variable domains greatly differ in among antibodies. V domain mediates antigen binding specificity of particular antibody for its variability is not evenly across the of the variable domains. Instead, the V regions of fragments called framework regions (FRs) of 15-30 acids by shorter regions of extreme variability called regions" or CDR. Each variable domain of native heavy and each comprise four FRs, largely adopting a beta-sheet connected by three hypervariable regions, which form loops and in some cases forming part of, the beta-sheet structure. hypervariable regions in each chain are held in proximity by the FRs and, with the hypervariable chain, contribute to the formation of antibodies. The constant domains are not an antibody to an antigen, but exhibit such as participation of the antibody in cytotoxicity (ADCC). The term "hypervariable region" refers to the amino acid residues of an for antigen binding. Typically, the comprises amino acid residues from a region" or and / or those residues a g In it may also be alter one or CDR amino in order to improve affinity to known "affinity “and be with for an to or and is not to affinity back vitro affinity maturation method by Wu et al., Proc Natl Acad Sci 95:6037 6042 (1998). "Framework regions" (FR) are those variable domain residues than the CDR residues. Each variable domain typically has four identified as FR1, FR2, FR3 and FR4. If the defined to Kabat, the light chain FR residues are about at 1-23 (LCFR1l), 35-49 (LCFR2), and 98-107 (LCFR4) and heavy chain FR residues are at residues 1-30 36-49 (HCFR2), 66-94 and (HCFR4) in heavy If the CDRs comprise light chain FR residues are 33-49 (LCFR2), 53-90 and 97-107 (LCFR4) the and the heavy chain are positioned about at (HCFR1), 33-52 (HCFR2), (HCFR3), and 102-113 (HCFR4) chain residues. In when the CDR from both a CDR and those of a the FR residues accordingly. For includes amino acids the heavy chain FR1 are positions 1-25 and the FR2 are at positions 36-49. The fragment region ("Fc region, Fc") of immunoglobulin is the of an immunoglobulin molecule interacts with cell Fc-receptor, as well as some proteins the complement system. property allows antibodies to activate immune system. In IgG, IgA and IgD antibody isotypes, the Fc is composed of two identical protein fragments, respectively, from second and third constant domains of the two heavy chains; in IgE isotypes, the Fc region contains three heavy chain constant (CH domains 2-4) in each polypeptide chain. An antibody of the present invention "which binds" antigen refers to an antibody capable of binding the sufficient affinity such that the antibody can be used as a and / or therapeutic agent targeting a protein or cell antigen, and slightly cross-reacts with other proteins. analytical methods: fluorescence-activated cell radioimmunoassay (RIA) or ELISA, in such embodiments, the antibody binding to a non-target protein is less than 10 % binding to a specific target protein. With regard to the an antibody to a target molecule, the term "specific "specifically binds to" or is "specific for" a particular or an epitope on a particular polypeptide target means is noticeably (measurably) different from a non-specific (for example, in the case of bH1-44 or DbH1-81, a interaction is binding to bovine serum albumin, casein, serum or neutravidin). Specific binding can be measured, for example, by binding of a molecule compared to binding of a control molecule. example, specific binding can be determined by competition with control molecule that is similar to the for of non-labeled target. In this case, the binding of the labeled target to least about 60 nM, or at least about 50 nM, or at least about or at least about 30 nM, or at least about 20 nM, or at least 10 nM, or at least about 8 nM, or at least about 6 or at about 4 nM, or at least about 2 nM, or at least about 1 or In one embodiment, the term "specific binding" refers to a molecule binds to a particular polypeptide or epitope on polypeptide without substantially binding to any other polypeptide epitope. The term "Ka" as used herein refers to the association (on) of a particular antibody-antigen interaction. The term "Kd" as used herein refers to the dissociation rate of a particular antibody-antigen interaction. "Binding affinity" generally refers to the strength of total of noncovalent interactions between a single binding molecule (e.g. an antibody) and its binding partner (e.g. an Unless indicated otherwise, "binding affinity" refers to (characteristic, true) binding affinity which reflects a interaction between members of a binding pair (e.g. antibody antigen). The affinity of a molecule X for its binding partner Y generally be represented by the dissociation constant (Kd). preferred Kd value is about 200 nM, 150 nM, 100 nM, 60 nM, 50 nM, 30 nM, 20 nM, 10 nM, 8 nM, 6 nM, 4 nM, 2 nM, 1 nM, or can be measured by common methods known in the art, described herein.Low-affinity antibodies are usually the antigen and tend to dissociate easily, whereas antibodies are usually faster at binding the antigen and longer in the bound state. A variety of methods of affinity are known in the art, any of which can be of the present invention. In one embodiment, "Kd" or "Kd value" is plasmon resonance assays using (BIAcore, Inc., Piscataway, N.J.) at chips at ~10 response units (RU). biosensor chips (CM5, BIZAcore Inc.) are dimethylaminopropyl) -carbodiimide hydroxysuccinimide (NHS) according to Antigen is diluted with 10 mM sodium into 5 (~0.2 uM) and then injected at a flow rate approximately 10 response units (RU) of injection of antigen, 1M ethanolamine unreacted groups. For kinetics measurements, two-fold of Fab (e.g. 0.78 nM to 500 nM) are injected in PBS 20 (PBST) at 25° C at a flow rate of approximately 25 (kon) and off-rates (koff) are calculated using a Langmuir binding model (BIAcore Evaluation Software simultaneous fitting the association and dissociation sensorgram. equilibrium dissociation constant (Kd) is calculated as the koff / kon. See, e.g. Chen, Y., et al., (1999) J. Mol. Biol. 293: 881. If the on rate exceeds 10% M! gs! by the surface assay above, then the on-rate can be determined by a quenching technique that measures increase or fluorescence emission intensity nm; 16 nm band-pass) at 25°C of a nM in PBS, pH in of spectrophometer (Aviv Instruments) or a 8000-series spectrophotometer (ThermoSpectronic) with a stirred cuvette. The term "koff" refers to the off rate constant of a interaction between a binding molecule and antigen. The off can be measured using bio-layer interferometry, Octet™ system. "kon" according to the present invention can be measured the above surface plasmon assays N.J.} Ing. ~10 RU) . BIAcore Inc.) are carbodiimide according to the 10 mM sodium acetate, pH 4.8, 5 (~0.2 and then at a flow rate of 5 ul / minute 10 units (RU) of coupled protein. ethanolamine solution is injected to unreacted groups. Unless specified otherwise, the term "biologically active" "biological activity" and "biological characteristics" with to a polypeptide of the invention means having the ability to a biological molecule. The term "biological molecule” refers to a nucleic a protein, a carbohydrate, a lipid, and combinations thereof. In embodiment, the biological molecule exists in nature. Antibody fragments, such as Fab and F(ab'}2 can prepared from whole antibodies using conventional papain or pepsin digestion of whole antibodies. Moreover, portions thereof and immunoadhesion molecules can be prepared standard recombinant DNA techniques, for example, as described The term "recombinant antibody" is intended refer to antibody that is expressed in a cell or cell line sequence (s) encoding antibodies, wherein said is not naturally associated with the cell. As used herein, the term "variant antibody" is intended to to an antibody which has an amino acid sequence which differs from amino acid sequence of a "parental" antibody thereof by adding, deleting and / or substituting one or more amino acid as compared to the sequence of a parental antibody. In a embodiment, a variant antibody comprises at least one or more one to twelve, e.g. two, three, four, five, six, seven, eight or ten, eleven or twelve; in some embodiments, a variant comprises from one to about ten) additions, deletions, substitutions of amino acids as compared to a parental antibody. some embodiments, such additions, deletions and / or substitutions made in the CDRs of a variant antibody. Identity or homology respect to the sequence of a variant antibody is defined herein as percentage of amino acid residues in the variant antibody that are identical antibody residues, after the sequences and if necessary, to maximum percent of sequence A variant antibody ability to bind the and preferably to an which the and in embodiments, to those of binding affinity, longer half-life, lower IC50 value or ability to inhibit antigen biological activity as to parental antibody. The variant antibody of particular is one which displays at least 2 times, (preferably at least 5 10 times or 20 times) enhancement in biological activity as to a parental antibody. The term "bispecific antibody" refers to an antigen-binding domain(s) that are capable of to distinct epitopes on a single biological or specific binding to epitopes on two distinct bispecific antibody is also referred to as specificity" or as being a "dual specificity" antibody. In a broad sense, the term "chimeric antibody" is intended refer to an antibody that comprises one or more regions of antibody, and one or more regions of one or several other typically, a partially human and partially non-human derived partially from a non-human animal, such as mice, like vermin, or the Camelidae such as llama and alpaca. antibodies are generally preferred over non-human antibodies to reduce the risk of a human anti-antibody immune response, human anti-mouse antibody immune response in the case of a antibody. An example of a typical chimeric antibody is one in the variable region sequences are murine sequences, while the region sequences are human. In the case of a chimeric antibody non-human portions may be subjected to further alteration in order humanize the antibody. The term "humanization” is intended to refer to the when an antibody has a fully or partially non-human example, a mouse or llama antibody obtained by lamas, respectively, with an antigen of interest, or is a antibody based on such an antibody of a mouse or llama, it is to substitute certain amino acids, in particular in the regions and constant domains of heavy and light chains, in avoid or minimize the immune response in humans. The the interaction of the antibody with the target antigen mainly in amino acid residues located in the six CDR heavy and light chain.For this reason, amino acid sequences CDRs are far more variable between individual antibodies than outside of CDRs. Because CDR sequences are responsible for antibody-antigen interactions, it is possible to express antibodies that mimic the properties of a specific naturally antibody, or more generally, of any specific antibody with acid sequence, e.g. by constructing expression vectors that CDR sequences from the specific antibody and framework sequences a different antibody. As a result, it is possible to "humanize" human antibody and, binding and affinity of the is not precisely predict the and the antibody of a typically more human origin can be humanized to reduce the risk of a human antibody response. For chimeric antibodies, humanization modification of the framework regions of variable Amino acid residues that are part of regions (CDRs) will be most often not humanization, although in some it be in order modify individual amino acid of a in to delete a glycosylation isomerization site, or or linked glycosylation is an chain to an asparagine in a sequence Asn-X-Thr, where X can any except Pro. Removal of an glycosylation site may mutating either the Asn Ser / Thr residue preferably by conservative asparagine and residues can occur on such as pH exposure. Asparagine are deamidation, primarily when present in sequence and lesser degree in other dipeptide sequences such a CDR sequence comprises such a deamidation site, Gly, it may be desirable to remove this site, conservative substitution to delete one of the residues. Numerous methods for humanization of an known in the art. One commonly used method grafting may be based on the CDR definitions by last edition (Magdelaine-Beuzelin et al., Crit 64:210 225 (2007)) suggests that the IMGT® ImMunoGeneTics information system®, improve humanization results (see Lefranc et al., Comp 27:55-77 (2003)). In some cases, CDR grafting may the specificity and affinity, and thus the biological of a grafted non-human antibody, as compared to a which the CDRs were obtained. Back mutations are referred to as "framework region repair") may be positions of a CDR grafted antibody, typically in order to restore the binding specificity and of a antibody. Identification of positions for possible be performed using information available in the antibody databases. Amino acid residues that are for mutations are typically those that are located at the surface of antibody molecule, whereas residues that are buried or that have a degree of surface exposure will not normally be altered. An humanization technique to CDR grafting and back resurfacing, in which non-surface exposed residues of are retained, whereas surface residues are altered to human Fully human antibodies can be generated using using in - collected phage libraries or in vivo humanized (mice, rats, etc.). of combinatorial phage with of a source of gene on library types can be and immune libraries which encode respectively. MRNA from the antibody-producing lymphoid cell isolated for this purpose. Peripheral blood lymphocytes are used, but in some cases splenocytes have been used as well Amersdorfer P, Finnern R, Sargent P, Lindquist E, Schier Efficient construction of a large the production of high-affinity human protein antigens. Proc Natl Acad Sci U S A and Haard HJ, van Neer N, Reurs A, Hufton SE, Roovers P, al. A large non-immunized human Fab rapid isolation and kinetic analysis Biol Chem 1999,274:18218-18230.], cells or lymphocytes [Vaughan TJ, Williams AJ, Pritchard K, Osbourn AR, Earnshaw JC, et al. Human antibodies with isolated from a large non-immunized phage Biotechnol 1996,14:309-314.]. cDNA is then on the mRNA, and both oligo-dT primers hexanucleotides can be used that yield cDNA variants of genes encoding the variable of AB, Kapralova MV, Laman AG, Shepelyakovskaya AO, Bulgakova EB, KK, et al. The library of human miniantibodies in the phage format: Designing and testing DAN: Izd-vo "Nauka"; 2005.]. One or several primers can be simultaneously used to limit range of amplified genes to one or several variable domain families or antibody isotypes, now at cDNA level [Marks HR, Bonnert TP, McCafferty J, Griffiths AD, Winter immunization. Human antibodies from V-gene libraries phage. J Mol Biol 1981,222:581-597]. The primers used for of genes encoding immunoglobulins are complementary to conservative regions. Their sequences are selected from collections that are organized into databases, such as Kabat or databases. The primer design also provides for internal sites for cloning the PCR-products into the appropriate vectors. Construction of synthetic libraries is based on replacement natural CDRs with a set of random sequences. In this case, it possible to generate a vast variety of antigen-binding sites. Phage display is one of the most powerful and widely used in technique for search for antibodies. In 1985, Smith found that DNA sequences could be cloned into filamentous bacteriophage that such cloned sequence can be expressed on the surface of particles as fusion proteins (Smith GP: Filamentous fusion phage: expression vectors that display cloned antigens on virion Science 1985, 228:1315-1317.). Thus, it is to fusion proteins of interest based on their proteins. This discovery was combined with PCR which made it possible to clone the cDNA genes to create a variety of phage domains that can be used to quickly monoclonal antibodies. Phage library - reflects of cell antibody of any human or animal was to the library. In 19395, two papers the mice which were of of repertoires of are to technology FA, Schramm Kuo comprising four distinct genetic modifications. Nature 1994, 859). In these animals, their own endogenous heavy and immunoglobulin chain genes were deliberately destroyed, introduction of transgenes, which are the segments of human heavy k light chain genes.It turned out human gene used by the mouse immune system to high affinity antibodies against a greater of fact that transgenic mice express are hybrids of mouse and human components (human Iga, IgB, and other signaling molecules), their mature normally. In certain cases, it may also be desirable to alter one or CDR amino acid residues in order to improve binding affinity to target epitope. This 1s known as "affinity maturation" and optionally be performed in connection with humanization, for in situations where humanization of an antibody leads binding specificity or affinity and it is not possible to improve the binding specificity or affinity by back Various affinity maturation methods are known in the in vitro scanning saturation mutagenesis et al., Proc Natl Acad Sci USA, 94:412-417 (1997) and the vitro affinity maturation method by Wu et al., Proc Natl 95:6037 6042 (1998). The term "monoclonal antibody" or "mAb" refers to an that is synthesized and isolated by a separate clonal cells. The clonal population can be a clonal population cells. In some embodiments, the immortalized cells in a population are hybrid cells -hybridomas - typically produced fusion of individual B lymphocytes from immunized animals individual cells from a lymphocytic tumour. Hybridomas are a type constructed cells and do not exist in nature. "Native antibodies" are usually heterotetrameric glycoproteins about 150000 daltons, composed of two identical light (L) chains two identical heavy (H) chains. Each light chain is linked chain by one covalent disulfide bond, while the of linkages varies among the heavy chains of different isotypes. Each heavy and light chain also has regularly intrachain disulfide bridges. Each heavy chain has at one end variable domain (VH) followed by a number of constant domains. light chain has a variable domain at one end (VL) and a at its other end. The constant domain of the light with the first constant domain of the heavy chain, variable domain is aligned with the variable domain of the Particular amino acid residues are believed to form an between the light chain and heavy chain variable domains. The term "isolated" used to describe various antibodies in description refers to an antibody which has been identified separated and / or regenerated from a cell or cell culture, in which antibody is expressed. Impurities (contaminant from natural environment are materials which diagnostic or therapeutic uses of and enzymes, or sequenator), or (2) to homogeneity by SDS-PAGE under reducing conditions using Coomassie Brilliant Blue, or silver stain. Isolated antibody includes the antibody in recombinant cells since at least one component of the natural environment will not be present. Isolated typically prepared by at least one purification An "isolated" nucleic acid molecule is is and separated from at least one nucleic acid the former is bound to in the natural source of An isolated nucleic acid molecule is different from in which it is found under natural conditions. Thus, an nucleic acid molecule is different from a nucleic acid exists in cells under natural conditions. An isolated molecule however includes a nucleic acid molecule located which the antibody is normally expressed, for example, if the acid molecule has a chromosomal localization that is different its localization in cells under natural conditions. The term "epitope" as used herein is intended to refer to portion (determinant) of an antigen that specifically binds to binding molecule (for example, an antibody or a related molecule, as a bispecific binding molecule). Epitope determinants consist of chemically active surface groupings of molecules amino acids or carbohydrates or sugar side chains and comprise specific three dimensional structural well as specific charge characteristics. Epitopes can "linear" or "conformational". In a linear epitope, all of of interaction between a protein (e.g. an antigen) and an molecule (such as an antibody) occur linearly along the acid sequence of the protein. In a conformational epitope, of interaction occur across amino acid residues the are separated from one another in the primary Once a desired epitope of an antigen is determined, generate antibodies to that epitope using techniques art. In addition, generation and characterization of other binding molecules may elucidate information about epitopes. Based on this information, it is then competitively screen antibodies for binding to the same or epitopes, e.g. by conducting competition studies to find molecules that compete with one another for binding to the The term "peptide linker" as used herein is intended to mean peptide having the ability to combine domains, with a length depends on the domains which it binds to each other, and any amino acid sequence. Preferably, the peptide linker has a of more than 5 amino acids and consists of any set of amino selected from G, A, 8S, P, E, T, D, K. The term "in vitro" refers to a biological entity, a process, or a biological reaction outside the body under conditions. For example, a cell grown in vitro is to be understood a cell grown in an environment outside the body, e.g. in a test a culture vial, or a microtiter plate. The term "ICsc" (inhibitory concentration 50%), as refers to concentrations of drug, at which a measurable response, for example, growth / proliferation of cells as cells, 1s inhibited by 50%. ICs; value can be calculated appropriate dose-response curves, using special statistical for curve fitting. The term GIS50 (growth inhibition 50%) refers concentrations of drug, at which proliferation of cells, such as cells, is inhibited by 50%. The term "ED50" (EC50) (50% effective dose / concentration) to concentrations of drug producing 50% biological effect (which include cytoxicity). The term "antiproliferative activity" is intended to refer stopping or inhibiting growth of cells, such as cancer The term antibody "effector function" refers to activities attributable to the Fc-region (native Fc-region or Fc-region amino acid variants) of an antibody, which antibody isotype. Examples of antibody effector functions binding and complement dependent cytotoxicity; Fc antibody-dependent cell-mediated cytotoxicity (ADCC); down regulation of cell surface receptors (e.g. B-cell and B-cell activation. "Antibody-dependent cellular cytotoxicity" or "ADCC" refers cell-mediated response, in which nonspecific cytotoxic cells express Fc receptors (FcR) (for example, natural killer (NK) neutrophils, and macrophages) recognize bound antibody on a cell and subsequently cause lysis or phagocytosis of the target The primary cells for mediating ADCC, NK cells, express FcyRJII whereas monocytes express FcyRI, FcyRII and FcyRIII. FcR on hematopoietic cells is summarized in Table 3 on page 464 and Kinet, Annu. Rev. Immunol 9: 457-392 (1391). To assess ADCC of a molecule of interest, an in vitro ADCC assay, such as described in U.S. Patent Nos. 5,500,362 or Useful effector cells for such assays mononuclear cells (PBMC) and Natural Killer (NK) or additionally, ADCC activity of the molecule of assessed in vivo, e.g. in an animal model such as that Clynes et al. PNAS (USA) 95: 652-656 (1998). "Human effector cells" are leukocytes which express one or FcRs and perform effector functions. Preferably, the cells express least FcyRIII and perform ADCC effector function. Examples of leukocytes which mediate ADCC include peripheral blood cells (PBMC), natural killer (NK) cells, monocytes, T and neutrophils; with PBMCs and NK cells being preferred. The cells may be isolated from a native source thereof, e.g. from or PBMCs as described herein. The terms "Fc receptor" or "FcR" are used to describe a that binds to the Fc region of an antibody. The native sequence human FcR. Moreover, a preferred one an IgG antibody (a gamma receptor) and includes receptors of the FcyRII, and FcyRIII subclasses, including allelic variants alternatively spliced forms of these receptors. FcyRII include FcyRIIA ("activating receptor") and FcyRIIB which have similar amino acid the domains thereof. contains tyrosine-based its Inhibiting receptor an inhibition motif 234 (1997)). FcRs are reviewed in Ravetch and Kinet, Annu. Rev. 9: 457-92 (1991). Other FcRs, including those to be identified future, are encompassed by the term "FcR" herein. The term includes the neonatal receptor, FcRn, which is responsible for transfer of maternal IgGs to the fetus. "Complement dependent cytotoxicity" and "CDC" refer to ability of a molecule to lyse a target in the presence The complement activation pathway is initiated by the first component of the complement system (Clg) to a molecule antibody) complexed with a cognate antigen. To assess activation, a CDC assay, e.g. as described in Gazzano-Santoro J. Immunol. Methods 202: 163 (1996). The term "identity" or "homology" is construed to mean percentage of amino acid residues in the candidate that identical with the residue of a corresponding which it compared, after comparing the sequences necessary to achieve the maximum percent sequence, and not considering any conservative as of the sequence identity. Neither N- insertions will be construed as reducing or and computer programs for the alignment are known Sequence identity may be measured using sequence (e.g. Sequence Analysis Software Package, Genetics Computer University of Wisconsin Biotechnology Center, 1710 University Madison, WI 53705). This software matches similar assigning a degree of homology to various substitutions, (eliminations), and other modifications. The term "homologous" with regard to a polypeptide sequence an antibody should be construed as an antibody exhibiting at 70%, preferably 80%, more preferably 90% and most preferably sequence identity relative to a polypeptide sequence. The term relation to a nucleic acid sequence should be construed as a of nucleotides exhibiting at least 85%, preferably 90%, 95% and most preferably 97% sequence identity relative to a acid of amino acid sequences of herein are For example, it may be desirable to binding and / or other biological properties of Amino sequence variants of antibody are prepared by nucleotide changes into the antibody nucleic synthesis. Such modifications include, for example, insertions and / or substitutions of residues within sequences of antibody. Any combination of deletion, and substitution is made to arrive at the final that the final construct possesses the desired amino acid changes also may alter post-translational antibody, such ‘as changing the number or position sites. Variant of modification of amino acid using amino acid substitutions. Such a variant least one amino acid residue in the antibody molecule a residue.The sites of interest for substitutional include hypervariable or CDRs, but FR or Fc are also substitutions are alteration of the biological activity, further substantial be made, which are denoted as "exemplary substitutions" set Table A, or alterations described in more detail below when amino acid classes, and also product screening may be performed. The terms "nucleic acid", sequence", "polynucleotide", sequence" and "nucleotide used present description, mean a sequence of or not, determining a fragment or a region a containing unnatural nucleotides or not, and either a strand DNA or RNA, a single-strand DNA or RNA, or products of said DNAs. It should also be included here that the present invention relate in their natural invention have been isolated and / or purified, i.e., they were directly or indirectly, for example by a copy, their environment been at least partially modified. Thus, isolated nucleic acids by recombinant genetics, by means, for example, of host cells, obtained by chemical synthesis should also be mentioned here. A reference to a nucleotide sequence encompasses the thereof unless otherwise specified. Thus, a reference to a acid having a particular sequence should be understood as one encompasses the complementary strand thereof with the sequence thereof. The term "control sequences" refers to DNA sequences for the expression of an operably linked coding sequence particular host organism. The control sequences that are suitable prokaryotes, for example, include a promoter, optionally an sequence, and a ribosome binding site. Eukaryotic cells are utilize promoters, polyadenylation signals, and enhancers. Nucleic acid is "operably linked" when it is placed into functional relationship with another nucleic acid sequence. DNA for a presequence or secretory leader sequence is DNA for a polypeptide if it is expressed as a in the secretion of the polypeptide; a operably linked to a coding sequence if it of the sequence; a ribosome binding site a coding sequence if it is positioned so as to Generally, "operably linked" means that the DNA are contiguous, and, in the case of a secretory and in reading phase. However, enhancers do not have to term "vector" as used herein means a nucleic acid capable of transporting another nucleic acid to which it linked. In some embodiments, a vector is a plasmid, i.e., a double stranded piece of DNA into which additional DNA be ligated. In some embodiments, a vector is a viral vector, additional DNA segments may be ligated into the viral genome. In embodiments, vectors are capable of autonomous replication in cell into which they are introduced (e.g. bacterial bacterial origin site of replication and episomal In further embodiments, vectors (e.g. non-episomal can be integrated into the genome of a host cell upon into a host cell, and thereby are replicated along with the Moreover, certain vectors are capable of directing the genes to which they are operatively linked. Such vectors are to herein as "recombinant expression vectors" (or simply, vectors"). The term "recombinant host cell" (or simply cell") herein is intended to refer to a cell into a expression vector has been introduced. The present to host cells, which may include, for example, a vector the invention described above. The present invention also host cells that comprise, for example, a nucleotide sequence a chain or antigen-binding portions thereof, a light nucleotide sequence or antigen-binding portions thereof, the first binding domain and / or second binding domain molecule of the invention. It be understood to a particular subject cell but to the progeny of such a cell well. Since modifications may occur in succeeding generations due either mutation or environmental influences, such progeny may fact, be identical to a parental cell, however, such cells included within the scope of the term "host cell" as used The term "excipient" is used herein to describe any other than the compound(s) of the invention. The term "disease or disorder mediated by GITR" refers disease or disorder that is either directly, or indirectly with GITR, including etiology, development, progression, or pathology of a disease or disorder. "Treat", "treating" and "treatment" refer to a method alleviating or abrogating a biological disorder and / or of attendant symptoms thereof. As used herein, to disease, disorder or condition means reducing the occurrence frequency of the symptoms of a disease, condition. Further, references herein to "treatment" references to curative, palliative and prophylactic treatment. In one aspect, the subject of treatment, or patient, is a preferably a human subject. Said subject may be either male or of any age. The term "disorder™ means any condition that would treatment with the compound of the present invention. The of this term includes chronic and acute disorders or diseases those pathological conditions that predispose the mammal to disorder in question. The terms "cancer" and "cancerous" refer to a condition or describe a physiological condition in typically characterized by unregulated growth / proliferation of The definition encompasses both benign and malignant diseases. Examples of cancerous diseases include, but are not to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. particular examples of such cancerous diseases include squamous cancer, small-cell lung cancer, non-small cell adenocarcinoma of the lung and squamous carcinoma of peritoneal cancer, hepatocellular cancer, stomach cancer gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, cancer, colon cancer, colorectal cancer, endometrial or carcinoma, salivary gland carcinoma, kidney or renal cancer, cancer, vulval cancer, thyroid cancer, hepatic carcinoma, carcinoma, penile carcinoma, melanoma, and various head and cancers. The terms "immune response", "autoimmune response" "autoimmune inflammation" refer, for example, to the action antigen-presenting cells, phagocytic cells, macromolecules produced by said cells or antibodies, cytokines and complement produced in the damage, destruction or elimination of invasive infected with pathogens, cancer cells or, of or inflammation, normal cells or from human effective amount" is intended to refer to amount of therapeutic agent being administered which will to some extent one or more of the symptoms of the disorder treated. The term "chronic" use refers to continued of agent (s) as opposed to acute (transient) route so as to sustain the initial therapeutic effect for a period of time. "Intermittent" use refers to treatment that is not carried consistently without interruptions, but which is rather periodic nature. As used herein, the words "comprise," "have, variations such as "comprises," "comprising," "has," "includes" or "including", and all grammatical variations thereof be understood to imply the inclusion of a stated integer or group integers but not the exclusion of any other integer or group integers. Detailed description of the invention Antibody The present invention relates to a monoclonal antibody specifically binds to GITR. In one aspect, the present invention relates to a antibody or antigen-binding fragment thereof that specifically to GITR comprising: (a) a heavy chain variable domain comprising: (1) CDR1 comprising an amino acid sequence selected from group: NYGMH (SEQ ID NO: 1) or YYWMY (SEQ ID NO: 12); (ii) CDR2 comprising an amino acid sequence selected from group: VIWFDGSNKFYTDSVKG (SEQ ID NO: 2) or AISWNGGRTYYAESMKG (SEQ 13); (iii) CDR3 comprising an amino acid sequence selected from group: ELGGYYYDSSGFRPYYYGMDV (SEQ ID NO: 3) or NRYYSDPNYGMNL (SEQ 14), and (b) a light chain variable domain comprising: (1) CDR1 comprising an amino acid sequence selected from group: RASQSIGSWLA (SEQ ID NO: 7) or TGTSTDIGTYKYIS (SEQ ID NO: 17); (ii) CDR2 comprising an amino acid sequence selected from group: AASTLOR (SEQ ID NO: 8) or GVSHRPS (SEQ ID NO: 18); (iii) CDR3 comprising an amino acid sequence selected from group: QQSHSHPLT (SEQ ID NO: 9) or SSYTSSGTVV (SEQ ID NO: In some embodiments, a monoclonal antibody or fragment thereof comprises a heavy chain variable CDR 1, 2 and 3 comprising amino acid sequences sequences of ID NOs: 1, 2 and 3, respectively. In some a monoclonal antibody or fragment thereof a heavy chain variable CDR 1, 2 and 3 amino acid sequences sequences of SEQ ID 13 and 14, respectively. In some embodiments, a monoclonal antibody or fragment thereof comprises a light chain variable CDR 1, 2 and 3 comprising amino acid sequences represented by NOs: 7, 8 and 9, respectively. In some embodiments, a monoclonal antibody or fragment thereof comprises a light chain variable CDR 1, 2 and 3 comprising amino acid sequences represented by NOs: 17, 18 and 19, respectively. In some embodiments, a monoclonal antibody or fragment thereof comprises: . a heavy chain variable domain comprising CDRs 1, 2 and comprising amino acid sequences represented by the sequences of ID NOs: 1, 2 and 3, respectively; a light chain variable domain comprising CDRs 1, 2 and comprising amino acid sequences represented by the sequences of ID NOs: 7, 8 and 9, respectively. In some embodiments, a monoclonal antibody or fragment thereof comprises: a heavy chain variable domain comprising CDRs 1, 2 and comprising amino acid sequences represented by the sequences of ID NOs: 12, 13 and 14, respectively; a light chain variable domain comprising CDRs 1, 2 and comprising amino acid sequences represented by the sequences of ID NOs: 17, 18 and 19, respectively. In some embodiments, a monoclonal antibody or fragment thereof comprises a heavy chain variable an amino acid sequence that is at least 90% homologous to acid ID NO: 4). In some embodiments, a monoclonal antibody or fragment thereof comprises a heavy chain variable the amino acid ID NO: 4). In some embodiments, a monoclonal antibody or fragment thereof comprises a heavy chain variable an amino acid sequence that is at least 90% homologous to acid (SEQ NO: 15). In some embodiments, a monoclonal antibody or fragment thereof comprises a heavy chain variable the amino acid (SEQ ID In some embodiments, a monoclonal antibody or fragment thereof comprises a light chain variable an amino acid sequence that is at least 90% homologous to acid sequence (SEQ ID NO: In some a monoclonal antibody or fragment thereof a light chain variable the acid (SEQ ID NO: In some a monoclonal antibody or fragment thereof a light chain variable an amino acid sequence that is at least 90% homologous to acid (SEQ ID NO: In some a antibody or fragment thereof a chain variable the acid (SEQ ID NO: In a monoclonal antibody or fragment CC -a heavy comprising an amino acid that 1s at to the amino acid ID NO: 4); - a light chain variable domain comprising an amino acid that 1s at least 90% to the amino acid (SEQ ID NO: In a monoclonal antibody or fragment -a domain amino ID NO: -a chain variable domain amino (SEQ ID NO: In some a monoclonal antibody or fragment thereof CC -a heavy domain comprising an amino acid that is at least to the amino acid sequence NO: (SEQ 15); - a light chain variable domain comprising an amino acid that 1s at least 90% to the amino acid (SEQ ID NO: In some a monoclonal antibody or fragment - domain amino TISRDNAQNTLYLQMNSLKSEDTAVYYCAKNRYYSDPNYGMNLWGKGTTVTIVSS (SEQ ID 15); - chain variable domain amino (SEQ ID NO: 20). In some a monoclonal antibody s full length IgG antibody. In some embodiments, the monoclonal IgG antibody is of human IgG2, IgG3 or IgG4 isotype. - In some embodiments, the monoclonal IgG antibody is of human isotype. In some embodiments, a GITR-specific monoclonal comprises the E345R mutation in the Fc fragment to increase properties, antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC). In some embodiments, a monoclonal comprising an amino acid sequence that at the (SEQ NO: In a monoclonal the amino (SEQ NO: In some a monoclonal comprising an sequence that at the (SEQ NO: In a monoclonal the amino (SEQ NO: In a monoclonal a the In a monoclonal antibody comprises a light the amino acid In some a monoclonal antibody comprises: - a comprising an amino acid sequence that is at 80% to the (SEQ - a an sequence that at 80% to the In some a monoclonal antibody that binds to GITR The comprises: a the amino acid (SEQ NO: a acid In a monoclonal antibody comprises: - a comprising an amino acid sequence that is at 80% the (SEQ NO: - a light ¢ n an amino that is at In some monoclonal antibody that binds to GITR The difference between BCD166-02-001 and BCD166-01-001 is E345R mutation in the Fc fragment to increase agonist antibody-dependent cellular cytotoxicity (ADCC), but not dependent cytotoxicity (CDC). The monoclonal antibody comprises: a chain the amino acid (SEQ NO: 5 acid In some a monoclonal antibody comprises comprising an sequence that is at least 90% homologous the (SEQ NO: a monoclonal amino (SEQ NO: In some a monoclonal comprising an sequence that at the TVEKTVAPTECS (SEQ ID NO: In embodiments, a monoclonal antibody comprises a light the amino acid TVEKTVAPTECS (SEQ ID NO: In some embodiments, a monoclonal antibody comprises: - a chain comprising an amino acid sequence that is at 80% to the (SEQ NO: 16). - a comprising an sequence that is at 80% to the TVEKTVAPTECS (SEQ ID NO: 21). In some embodiments, a monoclonal antibody that binds to GITR is BRCD166-01-014. The antibody comprises: a chain the amino acid (SEQ NO: 16). a the amino acid TVEKTVAPTECS (SEQ ID NO: 21). Nucleic acid molecules The present invention also relates to nucleic acid particular to sequences encoding a monoclonal specifically binds to GITR according to the invention, as herein, optionally including any peptide linker sequence, connected therewith. A reference to a nucleotide sequence encompasses the thereof unless otherwise specified. Thus, a reference to a acid having a particular sequence should be understood as one encompasses the complementary strand thereof with sequence thereof. The term "polynucleotide" as means polymeric form of either nucleotides that are at least length, or ribonucleotides, or deoxyribonucleotides or a of either type of nucleotide. The term includes single and stranded forms. In one aspect, the present invention to a nucleic molecule comprising a nucleotide sequence an amino sequence from ID NOs: 1-21. A acid molecule In one aspect, the present invention relates to comprising a nucleotide sequence encoding a monoclonal antigen-binding fragment thereof that specifically binds to GITR, comprises: (a) a heavy chain variable domain comprising: (1) CDR1 comprising an amino acid sequence selected from group: NYGMH (SEQ ID NO: 1) or YYWMY (SEQ ID NO: 12); (ii) CDR2 comprising an amino acid sequence selected from group: VIWFDGSNKFYTDSVKG (SEQ ID NO: 2) or AISWNGGRTYYAESMKG (SEQ 13); (iii) CDR3 comprising an amino acid sequence selected from group: ELGGYYYDSSGFRPYYYGMDV (SEQ ID NO: 3) or NRYYSDPNYGMNL (SEQ 14), and (b) a light chain variable domain comprising: (1) CDR1 comprising an amino acid sequence selected from group: RASQSIGSWLA (SEQ ID NO: 7) or TGTSTDIGTYKYIS (SEQ ID NO: 17); (ii) CDR2 comprising an amino acid sequence selected from group: AASTLOR (SEQ ID NO: 8) or GVSHRPS (SEQ ID NO: 18); (iii) CDR3 comprising an amino acid sequence selected from group: QQSHSHPLT (SEQ ID NO: 9) or SSYTSSGTVV (SEQ ID NO: 19). In some embodiments, a nucleic acid molecule nucleotide sequence that encodes monoclonal antibody or an binding fragment thereof comprising a heavy chain variable comprising CDRs 1, 2, and 3 comprising amino acid sequences by the sequences of SEQ ID NOs: 1, 2 and 3, respectively. In some embodiments, a nucleic acid molecule nucleotide sequence that encodes monoclonal antibody or an binding fragment thereof comprising a heavy chain variable comprising CDRs 1, 2, and 3 comprising amino acid sequences by the sequences of SEQ ID NOs: 12, 13 and 14, respectively. - In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or an fragment thereof comprising a light chain variable CDRs 1, 2, and 3 comprising amino acid sequences sequences of SEQ ID NOs: 7, 8 and 9, respectively. In some embodiments, a nucleic “acid molecule nucleotide sequence encoding monoclonal antibody or an fragment thereof comprising a light chain variable CDRs 1, 2, and 3 comprising amino acid sequences sequences of SEQ ID NOs: 17, 18 and 19, respectively. In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or fragment thereof comprising: heavy chain variable domain comprising CDRs 1, 2 and amino acid sequences represented by the sequences of 1D 3, respectively; chain variable domain comprising CDRs 1, 2 and acid represented by the sequences of In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or fragment thereof comprising: a heavy chain variable domain comprising CDRs 1, 2 comprising amino acid sequences represented by the sequences ID NOs: 12, 13 and 14, respectively; a light chain variable domain comprising CDRs 1, 2 comprising amino acid sequences represented by the sequences ID NOs: 17, 18 and 19, respectively. In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or an fragment thereof comprising a heavy chain variable an amino acid sequence that is at least 90% homologous to acid sequence of SEQ ID NO: 4. In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or an fragment thereof comprising a heavy chain variable the amino acid sequence of SEQ ID NO: 4. In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or an fragment thereof comprising a heavy chain variable an amino acid sequence that is at least 90% homologous to acid sequence of SEQ ID NO: 15. In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or an fragment thereof comprising a heavy chain variable the amino acid sequence of SEQ ID NO: 15. In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or an fragment thereof comprising a light chain variable an amino acid sequence that is at least 90% homologous to acid sequence of SEQ ID NO: 10. In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or an fragment thereof comprising a light chain variable the amino acid sequence of SEQ ID NO: 10. In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or an fragment thereof comprising a light chain variable an amino acid sequence that is at least 90% homologous to acid sequence of SEQ ID NO: 20. In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or an fragment thereof comprising a light chain variable the amino acid sequence of SEQ ID NO: 20. In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or fragment thereof comprising: - a heavy chain variable domain comprising an amino acid that is at least 90% homologous to the amino acid sequence NO: 4; - a light chain variable domain comprising an amino acid that is at least 90% homologous to the amino acid sequence NO: 10. In some a nucleic acid molecule nucleotide sequence monoclonal antibody or fragment thereof -a domain comprising the amino of SEQ ID 4; - a chain variable domain comprising the amino of SEQ ID 10. In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or fragment thereof comprising: - a heavy chain variable domain comprising an amino acid that is at least 90% homologous to the amino acid sequence of NO: 15; - a light chain variable domain comprising an amino acid that is at least 90% homologous to the amino acid sequence of NO: 20. In some embodiments, a nucleic acid molecule nucleotide sequence encoding monoclonal antibody or fragment thereof comprising: - a heavy chain variable domain comprising the amino of SEQ ID NO: 15; - a light chain variable domain comprising the amino of SEQ ID NO: 20. In some embodiments, the nucleic acid molecule nucleotide sequence encoding a monoclonal antibody that binds to GITR which is a full-length IgG antibody. In some embodiments, a nucleic acid molecule comprises nucleotide sequence encoding a monoclonal IgG antibody which is human IgGl, IgG2, IgG3 or IgG4 isotype. In some embodiments, a nucleic acid molecule comprises nucleotide encoding a monoclonal IgG antibody which is human IgGl In some a nucleic acid molecule nucleotide sequence a monoclonal that binds to GITR which the Fc increase agonist cellular (ADCCY., ‘but not In some a nucleotide sequence a a chain comprising an sequence that at to the sequence of SEQ ID NO: 5. In some embodiments, a nucleic acid molecule comprises nucleotide sequence encoding a monoclonal antibody comprising a chain comprising the amino acid sequence of SEQ ID NO: 5. In some embodiments, a nucleic acid molecule comprises nucleotide sequence encoding a monoclonal antibody comprising chain comprising an amino acid sequence that is at least 390% to the sequence of SEQ ID NO: 6. In some embodiments, a nucleic acid molecule nucleotide sequence encoding a monoclonal antibody a chain comprising the amino acid of SEQ ID NO: In some embodiments, a nucleotide sequence encoding a a chain comprising an amino acid sequence that at In some embodiments, a nucleic acid molecule nucleotide sequence encoding a monoclonal antibody a chain comprising the amino acid sequence of SEQ ID NO: In some embodiments, a nucleic acid molecule comprises nucleotide sequence encoding a monoclonal antibody that - a heavy chain comprising an amino acid sequence that at 90% homologous to the sequence of SEQ ID NO: 5; - a light chain comprising an amino acid sequence that is at 90% homologous to the sequence of SEQ ID NO: 11. In some embodiments, a nucleic acid molecule comprises nucleotide sequence encoding a monoclonal antibody that comprises: - a heavy chain comprising the amino acid sequence of SEQ ID 5; - a light chain comprising the amino acid sequence of SEQ ID TY In some embodiments, a nucleic acid molecule comprises nucleotide sequence encoding a monoclonal antibody that comprises: - a heavy chain comprising an amino acid sequence that is at 90% homologous to the sequence of SEQ ID NO: 6; - a light chain comprising an amino acid sequence that is at 90% homologous to the sequence of SEQ ID NO: 11. In some embodiments, a nucleic acid molecule comprises nucleotide sequence encoding a monoclonal antibody that comprises: - a heavy chain comprising the amino acid sequence of SEQ ID 6; - a light chain comprising the amino acid sequence of SEQ ID TY In some embodiments, a nucleic acid molecule comprises nucleotide sequence encoding a monoclonal antibody comprising a chain comprising an amino acid sequence that is at least 390% to the sequence of SEQ ID NO: 16. In some embodiments, a nucleic acid molecule nucleotide sequence encoding a monoclonal antibody a chain comprising the amino acid of SEQ ID NO: In some embodiments, a nucleotide sequence encoding a a chain comprising an amino acid sequence that at to the sequence of SEQ ID NO: 21. In some embodiments, a nucleic acid molecule nucleotide sequence encoding a monoclonal antibody a chain comprising the amino acid sequence of SEQ ID NO: In some embodiments, a nucleic acid molecule comprises nucleotide sequence encoding a monoclonal antibody that comprises: - a heavy chain comprising an amino acid sequence that is at 90% homologous to the sequence of SEQ ID NO: 16; - a light chain comprising an amino acid sequence that is at 90% homologous to the sequence of SEQ ID NO: 21. In some embodiments, a nucleic acid molecule comprises nucleotide sequence encoding a monoclonal antibody that comprises: - a heavy chain comprising the amino acid sequence of SEQ ID 16; - a light chain comprising the amino acid sequence of SEQ ID 3 In some embodiments, a nucleic acid is DNA. In any of the above embodiments, nucleic acid molecules can isolated. A nucleic acid molecule of the invention can be from source that produces a monoclonal antibody that GITR. In certain embodiments, a nucleic acid molecule of the can be synthesized, rather than isolated. In one embodiment of the invention, nucleic acid encoding VH (SEQ ID NO: 4 or SEQ ID NO: 15) or VL (SEQ ID NO: SEQ ID NO: 20) domains are transformed into antibody genes entire length by virtue of insertion into an expression vector encoding heavy chain constant (CH) or light chain constant domains, respectively, such that the VH segment is operatively to the CH segment (s) within the vector, and / or the VL operatively linked to the CL segment within the In embodiment of the invention, nucleic acid molecules VL domains are transformed into genes along the antibody by virtue of linking, e.g. ligating, a encoding VH and / or VL domains to a nucleic CH and / or CL domains using standard Nucleic acid molecules encoding the chains may then be expressed from a cell which introduced. Nucleic acid molecules may be used to express a large of a recombinant monoclonal antibody that specifically binds Vector In another aspect, the present invention relates to suitable for the expression of any of nucleotide sequences herein. The present invention relates to vectors comprising molecules that encode any of the amino acid of a antibody that specifically binds heavy chain sequences of a first light chain sequences of a The invention further relates to molecules encoding fusion proteins, fragments. In some embodiments, a monoclonal antibody that binds to GITR according to the invention is expressed by DNA partially or fully encoding the sequence of a first or binding domain (e.g. light and heavy chain sequences domain comprises light and heavy chain sequences), described above, in expression vectors such that the genes operatively linked to necessary expression control transcriptional and translational control sequences. vectors include plasmids, retroviruses, viruses (AAV), plant viruses, such as mosaic virus, cosmids, YACs, EBV and molecules may be ligated into a that translational control sequences vector serve function of regulating the and translation of the An control sequences may to host cell used. DNA or first and second domain comprises a heavy and light chain sequence) can be into individual vectors. In one embodiment, any combination DNA molecules 1s introduced into the same expression vector. molecules can be introduced into an expression vector by methods (e.g. ligation of complementary restriction sites on antibody gene fragment and vector, or blunt end ligation if restriction sites are present). A suitable vector is one that encodes functionally complete CH or CL immunoglobulin sequences, with appropriate restriction engineering so that any VH or VL sequence can easily be inserted expressed, as described above. HC- and LC-encoding of genes in vectors may contain intron sequences, resulting in enhanced antibody protein yields by stabilizing the corresponding intron sequences are flanked by splice donor and splice acceptor which determine where RNA splicing will occur. Location of sequences can be either in variable or constant regions of chains, or in both variable and constant regions when multiple are used. Polyadenylation and transcription termination may a native chromosomal site downstream of coding regions. A expression vector can also encode a signal peptide that secretion of an antibody chain from a host cell. An gene may be cloned into a vector such that the signal in-frame to the amino terminus of an immunoglobulin The peptide can be an immunoglobulin signal peptide or signal peptide (i.e., a signal peptide from a protein). In addition to antibody chain genes, the recombinant expression of the invention can carry regulatory sequences that the expression of antibody chain genes in a host «cell. will be understood by those skilled in the art that the design of expression vector, including the selection of regulatory may depend on such factors as the choice of a host to transformed, the level of expression of a desired and forth. Preferred control sequences for an expression host cell mammals include viral elements that ensure high levels of expression in mammalian cells, such as promoters and / or derived from a retroviral LTR, cytomegalovirus (CMV) (such as a promoter / enhancer), simian virus 40 (SV40) (such as a promoter / enhancer), adenovirus, (e.g. the major late adenovirus (AdMLP)), polyomavirus and strong mammalian as native immunoglobulin promoter or actin description of viral control elements and sequences US patents No. 5,168,062, 4,510,245 and 4,968,615. expressing binding molecules, such as antibodies in a description of promoters and vectors, as well as plants are known in the art. See, e.g. U. S. Patent Methods for expressing polypeptides in bacterial cells or e.g. yeast cells, are also well known in the art. In addition to antibody chain genes and regulatory recombinant expression vectors of the invention may carry sequences, such as sequences that regulate replication of a vector host cells (e.g. origins of replication) and selectable marker The selectable marker gene facilitates the selection of host into which a vector has been introduced (see U.S. Patent selectable marker gene confers resistance to medicinal agents, as G418, hygromycin or methotrexate, on a host cell into which a has been introduced. For example, selectable marker genes dihydrofolate reductase (DHFR) gene (for use in dhfr-host cells methotrexate selection / amplification), a neo gene (for selection), and a glutamate synthetase gene. The term "expression control sequence” as used herein is to refer to polynucleotide sequences that are necessary to effect expression and processing of coding sequences to which they ligated. Expression control sequences include transcription initiation, termination, promoter sequences; efficient RNA processing signals such as polyadenylation signals; sequences that stabilize sequences that enhance translation efficiency (i.e., sequence); sequences that enhance protein stability; and when sequences that enhance protein secretion. The nature of sequences differs depending upon the host organism; in such control sequences generally include the promoter of binding site, and transcription termination sequences; in typically, such control sequences include promoters and termination sequences. The term "control sequences" is include at least all components, the presence of which for expression and processing, and can also include components, the presence of which is advantageous, for example, sequences and fusion partner sequences. Host cells A further aspect of the present relates methods obtaining a monoclonal antibody to according to the invention. One a method for obtaining a that GITR, as defined herein, host cell capable of a binds to GITR, host cell monoclonal of a monoclonal binds to GITR. A antibody to by such in such host as "a monoclonal antibody that to GITR". The also relates to the progeny of from such host cells, monoclonal antibody that binds to GITR analogously. Nucleic acid molecules a monoclonal antibody specifically binds to GITR to the invention and comprising these nucleic acid be used for of a suitable mammalian or plant or may be introduced into mammalian cells by viral vectors. Methods transfecting cells are well known in the art. e.g. U.S. Pat. 4,740,461 and are well art, of and yeast cells are also well known art. Mammalian cell lines used as hosts for transformation are known in the art and include a plurality of immortalized available. These include, e.g. Chinese hamster cells, SP2 cells, HEK-293T cells, FreeStyle 293 NIH-3T3 cells, Hela cells, baby hamster kidney (BHK) green monkey kidney cells (COS), human hepatocellular (e.g. Hep G2), A549 cells, and a number of other cell Cell are selected by determining which cell lines have levels and provide for necessary characteristics of Other cell lines that may be used are insect cell or Sf21 «cells. When recombinant expression vectors monoclonal antibody that specifically binds to GITR are into mammalian host cells, the antibodies are produced by the host cells for a period of time sufficient to allow for of the antibodies in host cells or, more preferably, of antibodies into the culture medium in which the host cells are A monoclonal antibody that specifically binds to GITR can reconstituted from a culture medium using standard purification techniques. Plant host cells include, e.g. Arabidopsis, duckweed, corn, wheat, potato, etc. Bacterial host include Escherichia and Streptomyces species. Yeast host include Schizosaccharomyces pombe, cerevisiae and Pichia pastoris. Furthermore, level of production of specifically binds to GITR according to the cell line can be enhanced using a number of known example, the glutamine synthetase gene expression system (the system) is a common approach for enhancing expression under conditions. The GS system is discussed in whole or part in with EP No. 0216846, 0256055, 0323997 and 0338841. It is likely that a monoclonal antibody that specifically to GITR of various cell lines or transgenic animals will have different glycosylation profile as compared to each other. monoclonal antibody that specifically binds to GITR encoded by acid molecules described herein, or comprising amino acid provided herein are part of the present invention, regardless of glycosylation of the binding molecules, and, in general, of the presence or absence of post-translational modifications. Preparation of antibodies The invention also relates to methods and processes for a monoclonal antibody that specifically binds to GITR and binding fragments thereof. Monoclonal antibodies Monoclonal antibodies may be prepared using the hybridoma first described by Kohler, et al. Nature 256,1975, p. 495, or may prepared using recombinant DNA methods (US 4816567). In a hybridoma method, a mouse, or other host such as a hamster, is immunized according to the lymphocytes that produce or are capable of will specifically bind to protein used another embodiment, lymphocytes can be a immunization. After immunization, the lymphocytes are fused with myeloma cell line using a suitable fusing agent, such as glycol, to produce a hybridoma cell. The hybridoma cells, obtained in the above manner, may be in a suitable culture medium that preferably contains one or substances that inhibit the growth or survival of the parental myeloma cells. For if lack the enzyme hypoxanthine or HPRT), the culture medium hypoxanthine, aminopterin, and (HAT which prevent the growth of cells. Preferred cells, used as component myeloma cell those that fuse efficiently, support stable high level antibodies by the selected antibody-producing cells, are to a medium where the unfused parental cells are selected. myeloma cell lines are murine myeloma lines, such as those from MOPC-21 and MPC-11 mouse tumors available from the Salk Cell Distribution Center, San Diego, California, and SP-2 or Ag8-653 cells available from the American Type Rockville, Maryland, USA. Human myeloma and cell lines also have been described for the of antibodies (Kozbor, J. Immunol., 133, 1984, p. 3001). Preferably, the binding specificity of monoclonal produced by hybridoma cells is determined by by an in vitro binding assay, such as radioimmunoassay or linked immunosorbent assay (ELISA). The binding affinity of the monoclonal antibody can, for be determined by the Scatchard analysis described in Munson al., Anal. Biochem., 107:220 (1980). Once hybridoma cells that produce antibodies of the specificity, affinity, and / or activity are identified, the clones be subcloned by limiting dilution procedures by methods. Suitable culture media for this for D-MEM or RPMI-1640 medium. In addition, cells may grown in vivo as ascites tumors in an animal e.g, by (L.p.) injection of the cells into mice. The monoclonal antibodies secreted by the subclones can separated from the culture medium, ascites fluid, or serum conventional antibody purification techniques such as, for affinity chromatography (e.g. using protein A- or protein chromatography, hydroxylapatite dialysis, etc. the monoclonal antibodies is isolated conventional are capable of and chains of murine serve into expression vectors, which are then into host such as E. coli cells, simian COS cells, Hamster cells, or myeloma cells, that do not produce being transfected, to obtain the synthesis of in the recombinant host cells. In a further embodiment of the invention, monoclonal or antibody fragments can be isolated from antibody generated using the techniques described in al., Nature, 348:552-554 (1990). Clackson et al., Nature, (1991) and Marks et al., J. Mol. Biol., 222:581-597 (1991) the isolation of murine and human antibodies, respectively, libraries. Subsequent publications describe the production affinity (nM range) human antibodies by chain shuffling al., Bio / Technology, 10:779-783 (1992), as well as infection and in vivo recombination as a strategy for very large phage libraries (Waterhouse et al., Nucl. Res. 21:2265-2266 (1993). Thus, these techniques are alternatives to traditional monoclonal antibody hybridoma for isolation of monoclonal antibodies. DNA that encodes an antibody may be modified, for example, so to produce chimeric or fusion antibody polypeptides, for example, substituting heavy chain and light chain (CH and CL) constant sequences for the homologous murine sequences (US 4816567 et al., Proc. Natl. Acad. Sci. USA: 81:6851 (1984), or by fusing the immunoglobulin coding sequence with all or coding sequence of a non-immunoglobulin polypeptide polypeptide). The non-immunoglobulin polypeptide sequences can substituted for the constant regions of antibody, or they can substituted for the variable domains of antigen-binding antibody to create a chimeric bivalent antibody comprising one binding site having specificity for an antigen and another binding site having specificity for a different antigen. a range endogenous production. For has been that the homozygous deletion of heavy-chain region (JH) gene in chimeric and mice results complete inhibition of endogenous Transfer of the human germ-line immunoglobulin gene array germ-line mutant mice results in the production of human after antigen challenge 5545806, 5569825, 5591669 (all GenPharm); 5545807; and WO Alternatively, rhage al., Nature, 348:552-553 (1390) and antibody fragments in vitro (V) gene repertoire from immunized donor to technique, antibody V-region genes are cloned either major or minor coat protein gene of a filamentous as M13 or and displayed as functional antibody fragments on of a particle. Because filamentous particle a DNA of the genome, selections mimics some of B-cell properties. Phage display can be performed in variety of formats. Several sources of V-gene segments can be phage display. Clackson et al., Nature, 352:624-628 (1991) various arrays of anti-oxazolone antibodies from a small combinatorial library of V genes derived from the spleen of mice. A repertoire of V genes from unimmunized human donors constructed and antibodies against a diverse array (including self-antigens) can be isolated essentially techniques described by Marks et al., J. Mol. Biol. (1991) . As described above, human antibodies may also be generated by vitro activated B cells (see US 5567610 and 5229275). Antibody fragments In certain circumstances, it is advisable to use rather than whole antibodies. The small sizes the contributes to rapid clearance thereof and to better penetration into dense tumors. techniques have been developed for the production Traditionally, these fragments were derived of intact antibodies. However, these can now directly by recombinant host cells. Fab, ScFv can be expressed in and secreted from E. thus the production of fragments. can be isolated from libraries above. According to fragments can be isolated from E. to form F(ab") (Carter et al., (1992). According another approach, ) can isolated directly from recombinant host F(ab'), with increased in vivo half-life receptor residues are described in US 5869046. the production of antibody fragments will be apparent to in the art. In other embodiments of the invention, the choice is a single chain Fv fragment (scFv) (see WO 5571894 and US 5587458). Fv and scFv are the only species binding sites that are devoid of constant regions; as a result, are suitable for reduced nonspecific binding during in vivo fusion proteins may be constructed to yield fusion an protein at either N- or C-terminus of an scFv. The may also be a "linear antibody", e.g. as described Such linear antibody fragments may be monospecific or Pharmaceutical In another provides a composition a antibody that to GITR as an (or as the only A include at least one intended to improve, prevent, or treat disorders may with GITR. composition" means a composition monoclonal that specifically binds to GITR to invention and at components selected from consisting of acceptable and compatible such as solvents, auxiliary, agents, stabilizers, delivery controllers, the of the type and route of compositions of the present and of thereof will be undoubtedly apparent Pharmaceutical compositions should compliance with the GMP (Good The composition may comprise a stabilizers and solubilizers. a be achieved by agents slowing down of active ingredient, for example, aluminum of suitable carriers, solvents, agents water, ethanol, polyalcohols and and esters for injections. "Medicament (drug)" is a compound or a mixture of compounds as pharmaceutical composition in the form of tablets, capsules, lyophilisates, injections, infusion, ointments and other intended for restoration, improvement or modification functions in humans and animals, and for treatment and diseases, for diagnostics, anesthesia, contraception, others. Any method for administering peptides, proteins or which is accepted in the art may be suitably employed for antibody that specifically binds to GITR according to the The term "pharmaceutically acceptable" refers to one or compatible liquid or solid components that are suitable administration in a mammal, preferably a human. The term "excipient" is used herein to describe any other than the above ingredients of the invention. These are of inorganic or organic nature which are used in the manufacturing in order to give drug products physicochemical properties. The terms "buffer", "buffer composition”, "buffering refers to a solution, which is capable of resisting changes in the action of its acid-base conjugate components, and which allows drug of a monoclonal antibody that specifically binds to CD20 to changes in pH. Generally, the pharmaceutical composition has a pH in the range from 4.0 to 8.0. Examples of buffers but are not limited to, acetate, phosphate, citrate, succinate, etc. buffer solutions. The terms "tonic agent", "osmolyte" or "osmotic as to an excipient that can increase the a antibody formulation. "Isotonic" drug is a an pressure equivalent to that of human blood. have an osmotic pressure from about 250 to 350 agents used include, but are not limited to, and sucrose, amino acids, metal salts, for example, "Stabilizer" refers to an excipient or of two or excipients that provide the physical and / or stability of active agent. Stabilizers include amino for but not limited to, arginine, histidine, proline; surfactants, are not to, 20 (trade name: Tween 80 (trade name: polyethylene-polypropylene Poloxamer, Pluronic, sodium example, but are not limited to, acid, monothioglycerol, sulfurous for example, but are not limited to, (EDTA), diethylenetriaminepentaacetic (DTPA), etc. A pharmaceutical composition is "stable" if the retains physical stability and / or chemical activity thereof during the specified shelf at temperature, for example, of 2-8 °C. Preferably, the active retains both physical and chemical stability, as well as activity. Storage period is adjusted based on the results of test in accelerated or natural aging conditions. A pharmaceutical composition of the invention can manufactured, packaged, or widely sold in the form of a single dose or a plurality of single unit doses in the form of a formulation. The term "single unit dose" as used herein discrete quantity of a pharmaceutical composition predetermined quantity of an active ingredient. The active ingredient typically equals the dose of the to be administered in a subject, or a convenient for example, half or a third of such Pharmaceutical compositions to the present are typically suitable for administration as formulations intended for in human breach in skin or the tract by virtue For or can also be employed. also Prefered embodiments of the subcutaneous routes. Any or proteins, which is accepted in be suitably for a monoclonal antibody that to GITR the invention. may prepared, or such as in plastic Formulations suspensions, or aqueous and the like. prior to Such formulation may be prepared example, process, which is known in the art as drying, freezing a product followed by removal solvent from frozen A monoclonal that to GITR to the invention can be inhalation, either alone, as a mixture with a acceptable excipient from an inhaler, such as a container, pump, spray, atomiser, or nebuliser, a propellant is used or not used, or as nasal drops, or Dosage forms for parenteral administration may be be immediate or modified release. Modified release include delayed-, sustained-, pulsed-, controlled-, targeted programmed release. Therapeutic use of a monoclonal antibody that specifically to GITR according to the invention In one aspect, a monoclonal antibody that specifically GITR according to the invention is useful in the treatment of that are associated with (mediated by) GITR activity. In one aspect, the subject of treatment, or patient, is a preferably a human subject. Said subject may be either male or of any age. In some embodiments, said monoclonal or fragment thereof that specifically binds to GITR a disease or disorder mediated by GITR, where the or is selected from the group comprising: cervical cancer, head and cancer, stomach cancer, breast cancer, renal cell cancer, (colorectal cancer), (OC) ovarian cancer, NSCLC (non-small cell cancer). In the case of a tumor (for example, cancer), the effective amount of an antibody or fragment thereof (for example, antibody or fragment thereof that specifically binds to GITR reduce the number of cancer cells; reduce the initial tumor inhibit (i.e., slow to some extent and preferably stop) cancer infiltration into peripheral organs; inhibit (i.e., slow to some and preferably stop) tumor metastasis; inhibit to some extent growth; and / or relieve to some extent one or more of associated with the disorder. The antibody or fragment some extent prevent growth and / or kill existing cancer cells, be cytostatic and / or cytotoxic. For cancer therapy, in vivo can, for example, be measured by assessing survival, time to progression (ITP), tumor response rate to treatment duration of response and / or quality of life. As used herein, the terms "co-administration”, and "in combination with", referring to a monoclonal specifically binds to GITR and one or more different agents, are intended to mean, refer to or include the 1) simultaneous administration of such of monoclonal antibody that specifically binds to GITR to invention and therapeutic agent to a patient in need of when such components are formulated together into a single dosage which releases said components at substantially the same time to patient, 2) simultaneous administration of such combination of monoclonal antibody that specifically binds to GITR according to invention and therapeutic agent to a patient in need when such components are formulated apart from each other dosage forms which are taken at substantially the patient, whereupon said components are released at same time to said patient, 3) sequential administration of such combination of a antibody that specifically binds to GITR according to the and therapeutic agent to a patient in need of treatment, when components are formulated apart from each other into separate forms which are taken at consecutive times by said patient significant time interval between each administration, whereupon components are released at substantially different times to patient; and 4) sequential administration of such combination of a antibody that specifically binds to GITR according to the and therapeutic agent to a patient in need of treatment, when components are formulated together into a single dosage form releases said in a controlled manner, whereupon they concurrently, or jointly released at the different times to where each portion may be by either the same or routes. A monoclonal that specifically binds to GITR to the invention can administered without further treatment, i.e., as an Furthermore, a monoclonal antibody that to invention may comprise at one (combination therapy). In of monoclonal antibody that may be jointly or formulated for treatment of a cancer. The term "cytotoxic agent" as used herein refers to a that inhibits or prevents the function of cells and / or destruction of cells. The term is intended to include isotopes (e.g. At2ll, TI131 T125 YS, Rel8€, Rel, gmls3, radioactive isotopes of Lu), chemotherapeutic agents, and as small molecule toxins or enzymatically active toxins of fungal, plant or animal origin, including fragments and / or thereof. "chemotherapeutic agent" is a chemical compound used a tumor. Examples of chemotherapeutic agents agents such as thiotepa and cyclosphosphamide such as busulfan, improsulfan and as benzodopa, carboquone, methylamelamines acid; teniposide; cryptophycins (e.g. 8); dolastatin; duocarmycin (including 218% and CB1-TM1); eleutherobin; spongistatin; nitrogen such cholophosphamide, mechlorethamine phenesterine, such as as the gamma II and Ed. Engl., A; an we TiC such as (e.g. T-2 verracurin dacarbazine; mannomustine; gacytosine; arabinoside ("Ara-C"); (TAXOL®), albumin-engineered (ABRAXANETM), and docetaxel mercaptopurine; methotrexate; platinum oxaliplatin, and carboplatin; vincas, polymerization fromforming microtubules, (VELBAN®), vincristine (ONCOVIN®),vindesine vinorelbine (NAVELBINE®) ; etoposide leucovorin; difluorometlhylornithine (DMFO); retinoidssuch as including bexarotene (TARGRETIN®) ; bisphosphonates (for example, BONEFOS® or OSTAC®), etidronate NE- zoledronic acid / zoledronate (ZOMETA®) , pamidronate (AREDIA®), tiludronate (SKELID®), or troxacitabine (1,3-dioxolane nucleoside oligonucleotides, particularly those that in signaling pathways implicated in aberrant as for example, PKC-alpha, Raf, H-Ras, and growth receptor (EGF-R); vaccines such as THERATOPE® and vaccines, for example, ALLOVECTIN® vaccine, LEUVECTIN® VAXID® vaccine; topoisomerase ~ inhibitor (e.g. ABARELIX®); BAY439006 (sorafenib; perifosine, COX-2 inhibitor (e.g. celecoxib or inhibitor (e.g. PS341); bortezomib (VELCADE®); 1577); orafenib, ABTS510; Bcl-2inhibitor such as (GENASENSE®) ; pixantrone; EGFR inhibitors (see tyrosine kinase inhibitors (see definition below); pharmaceutically acceptable salts, acids or derivatives of any of above; as well as combinations of two or more of the above such CHOP, an abbreviation for a combined of doxorubicin, vincristine, and abbreviation for a treatment regimen with combined with 5-FU and leucovovin. Also included in this definition are anti-hormonal agents act to regulate or inhibit hormone action on tumors, such as estrogens with mixed agonist / antagonist profile, including, (NOLVADEX®) , 4-hydroxytamoxifen, toremifene (FARESTON®), droloxifene, raloxifene (EVTSTA®) , trioxifene, selective estrogen receptor modulators (SERMs), such as anti-estrogens without agonist properties, such as (FASLODEX®) , and EM800 (such agents may block receptor dimerization, inhibit DNA binding, increase ER suppress ER levels); aromatase “inhibitors, aromatase inhibitors, such as formestane and exemestane and nonsteroidal aromatase inhibitors, such as (AREVIIDEX®), letrozole (FEMARA®) and aminoglutethimide, and aromatase inhibitors including vorozole (RIVISOR®), megestrol (MEGASE®) , fadrozole, imidazole; lutenizing hormone-releasing agonists, including leuprolide (LUPRON® ELIGARD®), buserelin, and tripterelin; sex steroids, as megestrol acetate and medroxyprogesterone as diethylstilbestrol and premarin, and fluoxymesterone, all transretionic acid anti-progesterones; estrogen receptor androgens, such as flutamide, and testolactone; and pharmaceutically salts, derivatives of any of the above; as well as combinations of two more of the above. Other therapeutic agents that can be used in with antibody that specifically binds to GITR to the inhibitors of growth example, include growth factor factor antibody cetuximab [Erbitux, C225] and any growth factor receptor antibodies disclosed by Stern et al. in oncology / haematology, 2005, Vol. 54, ppll-29); agents such as those which inhibit the effects of vascular growth factor, [for example, the anti-vascular endothelial factor antibody bevacizumab (Avastin)], anti-vascular growth factor receptor antibodies, such as anti-KDR anti-fltl antibodies; antisense therapies, for example directed to the targets listed above, such as ISIS 2503, an antisense or G3139 (Genasense), an anti bcl2 antisense; gene approaches, including, for example, approaches to replace genes, such as aberrant p53 or aberrant BRCAl or BRCA2, GDEPT directed enzyme pro-drug therapy), approaches such cytosine deaminase, thymidine kinase or a bacterial enzyme and approaches to increase patient tolerance to or radiotherapy, such as multi-drug resistance gene immunotherapy approaches, including, for example, treatment Alemtuzumab (campath-1H), a monoclonal antibody directed at CD52, treatment with antibodies directed at CD22, ex vivo and in approaches to increase the immunogenicity of patient tumour transfection with cytokines such as interleukin 2, interleukin 4 granulocyte macrophage colony stimulating decrease T cell anergy, such as treatment with inhibiting CTLA-4 function, approaches using transfected such as cytokine transfected dendritic cells, cytokine transfected tumour cell lines and idiotypic antibodies, adoptive T cells transfer have been non-specifically activated or targeted to a of interest ex vivo; inhibitors of protein such proteasome inhibitor, such as Velcade therapeutic approaches, for example, use proteins (such as antibodies or soluble external constructions), which either sequester receptor ligands, binding to receptor or decrease receptor signalling (for example, to enhanced receptor degradation or lowered expression levels). Other therapeutic agent that can be used in combination with antibody that specifically binds to GITR according to the can be an antibody selected from the group comprising: antibodies, anti-PD-L1 antibodies, anti-CTLA4 antibodies, antibodies, anti-0X40 antibodies or combinations Other therapeutic agent that can be used in antibody that specifically binds to GITR according to can be a therapeutically active antitumour compound selected from group of activators of innate or adaptive immunity. It is meant that a monoclonal antibody that to GITR according the invention may be used in the as described above, may be used in the treatment as and / or may be used in ‘the manufacture of a medication for as described above. Doses and routes of A monoclonal antibody binds to GITR to the invention will be an amount that is in of i.e. in and therapeutically effective amount may vary according to as the particular condition being treated, the age, sex the patient, and whether the monoclonal antibody that binds to GITR is being administered as a stand-alone or combination with one or more additional Dosage regimens may be adjusted For example, a single bolus may be may be administered over time or the or increased as indicated by the the situation. It is especially advantageous to compositions in a unit dosage form for ease uniformity of dosage. A unit dosage form as used herein to refer to physically discrete units suited as unitary patients / subjects to be treated; each unit contains a quantity of active compound calculated to therapeutic effect in association with the carrier. Specification for the unit dosage forms typically dictated by and directly dependent on (a) the characteristics of a chemotherapeutic agent and particular or prophylactic effect to be achieved, and (b) the limitations in the art of compounding such an active compound for the of sensitivity in the subjects. Thus, a skilled artisan would based disclosure provided herein, that the doses and adjusted in accordance with methods well-known in That is, the maximum tolerable dose can be the effective amount providing a detectable patient may also be determined, as can the administering each agent to provide a to a patient. Thus, while certain dose and exemplified herein, these examples in no way the administration regimen that may be provided to a patient in the embodiments of the invention. It is to be noted that dosage values may vary with the type severity of the condition to be alleviated, and may include single multiple doses. Furthermore, it is to be understood that for particular subject, specific dosage regimens should be adjusted time according to the individual need and the judgment of a professional administering or supervising the administration compositions, and that dosage ranges set forth herein are only and are not intended to limit the scope or practice of compositions. Further, the dosage regimen with the this invention may be based on a variety of factors, type of disease, the age, weight, sex, medical patient, the severity of the condition, the and the particular monoclonal antibody that to employed. Thus, the dosage regimen can vary can determined routinely using standard methods. For doses be adjusted based on pharmacokinetic or which may include clinical effects values. Thus, the encompasses the person skilled in art. and regimens known the a skilled Examples of suitable administration methods It is believed that a suitable dose of a specifically binds to GITR according to the be range of 0.1-200 mg / kg, preferably 0.1-100 mg / kg, 50 mg / kg, for example about 1-20 mg / kg. A monoclonal specifically binds to GITR may be administered, e.g. in a least 0.25 mg / kg, such as at least 0.5 mg / kg, including at least mg / kg, e.g. at least 1.5 mg / kg, such as at least 2 mg / kg, least 3 mg / kg, including at least 4 mg / kg, e.g. at least 5 for example up to a maximum of 50 mg / kg, including up to a 30 mg / kg, e.g. up to a maximum of 20 mg / kg, including up to a of 15 mg / kg. The administration will typically be repeated appropriate time intervals, such as once a week, once every two once every three weeks or once every four weeks, and for as long deemed appropriate by a responsible physician, who may, in some increase or reduce the dose if necessary. Diagnostic use and compositions A monoclonal antibody that specifically binds to GITR to the invention is also used in diagnostic processes (e.g. ex vivo). For example, the present monoclonal antibody specifically binds to GITR according to the invention can be used detecting or measuring the level of GITR in samples obtained from patient (e.g. tissue sample or a sample of body fluid, such as inflammatory exudate, blood, serum, intestinal fluid, saliva urine). Suitable methods for detection and immunoassays, such as flow cytometry, assay (ELISA), chemiluminescent assay, radioimmunoassay, immunohistology. The invention further includes kits, for diagnostic kits comprising a monoclonal antibody that binds to GITR described herein. Examples The following examples are provided for better understanding the invention. These examples are for purposes of illustration and are not to be construed as limiting the scope of the invention any manner. All publications, patents, and patent applications cited in specification are incorporated herein by reference. Although foregoing invention has been described in some detail by way illustration and example for purposes of clarity of will be readily apparent to those of ordinary skill in the art of the teachings of this invention that certain changes modifications may be made thereto without departing from the and scope of the appended embodiments. Materials and general methods General information regarding the sequences of immunoglobulin light and chains is Kabat, al., Sequences of Proteins of ed., Health Service, National Health, MD Amino acids of antibody are numbered to 63 (1969) 78-85; Kabat, E.A., et al., Sequences of Immunological Interest, 5th ed., Public Health Service, Institutes of Health, Bethesda, MD, (1991). Recombinant DNA techniques Standard methods were used to manipulate DNA as in Sambrook, J. et al, Molecular cloning: A laboratory manual; Spring Harbor Laboratory Press, Cold Spring Harbor, New York, The molecular biological reagents were used according to manufacturer protocols. Gene synthesis Desired gene segments were prepared from oligonucleotides by chemical synthesis. The gene segments of 300-4000 kb long, were flanked by singular restriction sites, were and ligation of oligonucleotides including PCR subsequently cloned via the indicated restriction sequences of the subcloned gene fragments were by sequencing. DNA sequence determination DNA sequences were determined by Sanger sequencing. DNA and protein sequence analysis and sequence The Infomax's Vector NT1 Advance suite version was sequence creation, mapping, analysis, annotation and Expression vectors For the expression of the described antibodies and variants of expression plasmids intended for expression in cells (E.coli), transient expression in eukaryotic cells cells) were applied. Beside the antibody expression vectors comprised: an origin of replication which allows of said plasmid in E. coli, genes which confer resistance E. to various antibiotics (e.g. to ampicillin and kanamycin). The fusion genes comprising the described antibody chains described below were generated by PCR and / or gene synthesis assembled with known recombinant methods and techniques by of the according nucleic acid segments, e.g. using unique sites in the corresponding vectors. The subcloned sequences were verified by DNA sequencing. For transfections, larger quantities of the plasmids were plasmid preparation from transformed E. coli cultures. Example 1 Production of recombinant antigens and antibodies in culture of mammalian To prepare antigens based on the sequence extracellular of GITR we generated number of domain GITR Macaca Mus from GITR gene sequence was for Fc IgGl Lama glama-tagged (Fig. at restriction sites. Furthermore, is separated from Fc by protease labile site of TEV protease; the was also used to cleave Fc fragment by treatment with TEV protease to obtain a tagless version of antigen. The sequence was also into plasmid containing FLAG-EPEA-tag (C-tag GE Healthcare) terminus of protein (Fig. 2) at SalIl / Notl restriction Furthermore, recombinant human GITR-ligand https: / / www.uniprot.org / uniprot / Q9%UNG2) was cloned in fashion into plasmid containing homotrimeric foldon [1] EPEA-tag at C-terminus of GITRL (Fig. 3). The required plasmids were produced in E.Coli cells and purified Qiagen kit. Antibodies and antigens were generated in established cell obtained from Chinese hamster ovary cells (CHO-K1) published protocols [Biotechnol Bioeng. 2005 Sep 20; Liao Metal., 2004; Biotechnol Lett. 2006 Biotechnol Bioeng. 2003 Nov 5;84(3):332-342]. expressing the EBNAl (Epstein-Barrvirus nuclear 1 gene were used. Suspension culture was conducted in on shaker using serum-free media from Life Technologies in accordance with manufacturer’s guidelines. For expression, cells at a concentration of 2*10% / ml were means of linear polyethyleneimine (PEI MAX, Polysciences). ratio was 1:3-1:10. In 5-7 days after transfection, cell centrifuged at 2000 g for 20 min and filtered through 0.22 um Target proteins were isolated from culture liquid by affine HPLC. Recombinant GITR and GITRL proteins comprising an (glutamic acid-proline-glutamic acid-alanine) at protein were isolated and purified from culture liquid using CaptureSelect tag Affinity Matrix sorbent. The culture liquid was passed chromatographic column pre-filled with 5 ml of C-tag column was then washed with 25 ml of PRS to remove binding components. Bound antigen was eluted under using 20mM Tris, 2M MgCl2 pH7.0-7.4. Then protein was into PBS (pH 7.4) using a semi-permeable dialysis membrane, (0.22 um), transferred into tubes and stored at -70°C. Recombinant GITR-TEV-Fc antigen was isolated and purified culture liquid on Protein A column for affine HPLC. Cleared liquid was passed through 5 ml HiTrap rProtein A Sepharose FF (GE Healthcare) equilibrated with phosphate buffered saline 7.4). Then, the column was washed with 5 column volumes remove non-specifically binding Bound antigen was using 0.1 M glycine buffer (pH 3). protein elution was collected and brought to neutral with M Tris All stages were conducted 110 flow rate. was dialyzed into PBS Dialysis filtered (0.22 um), tubes and stored at All recombinant were purified on a evaluated by SDS gel electrophoresis (examples are shown in Figs. and 5). Example 2 Construction of a naive human antibody Fab-library MeganLibTM Total RNA of B lymphocytes from blood samples from more than thousand individual human donors was isolated using RNeasy Mini according to the suggested protocol (QIAGEN). RNA concentration was performed using Nanovue kit (GE Healthcare), the quality isolated RNA was tested by means of 1.5% agarose gel Reverse transcription reaction was conducted RT (Evrogen) according to the recommended protocol transcriptase and random hexamer oligonucleotides as Reverse transcription products were used as a a stage polymerase chain reaction to obtain variable by restriction sites; reaction was performed using according to protocols by [J Biol Chem. 199% Jun 30]. The resulting DNA preparation VL-CK-VH (Fig. 6) was NheI / Eco911 restriction endonucleases and ligated into phagemid pHS (Fig. 7). Ligation products were transformed strain electrocompetent cells prepared in accordance with [Methods Enzymol. 2000;328: 333-63.]. Repertoire of phage Fab display library MeganLibTM was 101! transformants. library phage products were prepared in accordance with the described procedure [J Mol Biol. 1991 Dec 5;222(3): 581-97]. Example 3 Construction of immune Fab libraries of hybrid antibodies (CHARM). Total RNA of B lymphocytes from individual blood samples Glama immunized with recombinant GITR antigens, which maximum specific serum titer, was isolated using RNeasy according to the suggested protocol (QIAGEN). RNA concentration was performed using Nanovue kit (GE Healthcare), isolated RNA was tested by means of 1.5% agarose gel Reverse transcription reaction was conducted RT (Evrogen) according to the recommended protocol transcriptase and random hexamer oligonucleotides as Reverse transcription products were used as a a stage polymerase chain reaction to obtain variable by restriction sites; reaction was performed using according to protocols by [J Biol Chem. 1999 Jun 25; 30]. The resulting DNA preparation VL-CK-VH (Fig. 6) was NhelI / Eco911 restriction endonucleases and ligated into the phagemid pHS (Fig. 7). Ligation products were transformed into strain electrocompetent cells prepared in accordance with [Methods Enzymol. 2000;328: 333-63.]. Repertoire of phage Fab display library MeganLibTM was 1011 library phage products were prepared in accordance the described procedure [J Mol Biol. 1991 Dec 5;222(3): 581-97]. Table shows summary table of immune library products. Example 4 Selection of phage antibody Fab-libraries Specific anti-GITR human phage Fab antibodies were obtained the combinatorial phage Fab display library MeganLibTM and immune libraries described in Examples 2 and 3. Selection recombinant human GITR antigens by phage display [Nat Mar;14(3):309-14; J Mol Biol.1991 Dec 5;222(3): 581-97]. Selection of immune phage libraries was performed on EIA / RIA High Binding 5 ml immunotube. Antigen (0.5 ml 2 ug / ml) was sorbed carbonate buffer (0.1 M NaHCO3, pH 9.5) overnight at + 4C. Tubes then washed with PBST (10 mM phosphate buffered saline pH 7.3-7.5, mM NaCl and 2.7 mM KCl, 0.1% Polysorbate 20), and total volume (5 of 0.5% milk powder in PBST was added for blocking. The blocking alternated from round to round; for example, if 0.5% milk in PBST used in the 1st round, then 1% BSA in PBST was used in the 2nd Tubes were incubated for I h at room temperature on a shaker. were washed with PBST, and 2 ml / tube of blocking buffer and libraries were added thereto to a concentration of about 2*1012 particles per ml. We incubated them with phages for 1-2 hours temperature. In further rounds, we used 4 ml of phage supernatant a previous round, which was pre-clarified in a centrifuge 17,000 for 10-15 minutes. Tubes were washed 20 times with PBST. eluted from surface of tubes with 0.5 ml of 0.1 M glycine 2.2: glycine buffer was added to the tubes, and the lightly stirred for 15 minutes at room temperature. Phage then transferred to clean, non-sorbing tubes with 100 pl of 1M pH8.0 for neutralization. Tubes were kept in ice until the cells infected. After selection, M13 bacteriophage is cultured using E.coli strain as a host. Amplification is carried out by infecting the strain culture with phages followed by growing for 12-15 hours. After selection, 0.5-1 ml of phages was mixed with a cell (ODeco=0.3-0.4) and incubated for 1.5 hours at 37°C. Then, cells spun down at 3,000-4,000 rpm for 10-15 minutes, resuspended in : of medium and plated on Petri dishes containing an antibiotic selection (ampicillin). Colonies were grown in a thermostat at After 12-15 hours, we calculated the number of colonies and cells washed from dishes with 5-10 ml of LB medium. 100 pl of cell was added to 20 ml of antibiotic medium (ampicillin). We density to 0D600=0.35-0.5 by 37. K07 phage helper was added at 10 ml of culture (10!% particles / ml) and incubated at 37C for 1.5 without shaking. Then, an equal volume of medium with ampicillin (100 pg / ml) and double-dose kanamycin (40 pg / ml) dose IPTG (0.2 mM) was added to the cell culture. The cell culture loaded into a shaker, and phages were cultured for 4-5 hours at The cell culture was centrifuged for 25-30 min at 17,000 g, was collected in tubes. The resulting supernatant was then used next round of selection, or phage was isolated by precipitation further storage. Phages solution was added supernatant The incubated ice for at least 3 hours. The solution centrifuged for 10 minutes at 8,000 g, the resulting comprising phages was diluted in °~ ml of TBS buffer batfer). After 2-3 rounds, bacteriophages were isolated, we specific binding of polyclonal phage to human GITR, and antigens. Example 5 Analysis of polyclonal phages from 2nd and 3rd rounds selection. After 3 rounds of selection of 2 naive MeganLib libraries and immune libraries on target antigen, we examined specific and specific binding of polyclonal phages from 2nd and 3rd rounds ELISA. The target GITR antigen was sorbed overnight in a (0.1 M NaHCO3, pH 9.5) onto the plastic surface of the ELISA plate analyze for the presence of specifically binding phages (2 ug / ml), GCSF, hIL6R-Fc, interferon alfa-2b, Rituximab (2 ug / ml) to analyze non-specifically binding phages. Plates were then washed with PBST mM phosphate buffered saline pH 7.3-7.5, 137 mM NaCl and 2.7 mM 0.1% Polysorbate 20), then 300 pl / well of 0.5% milk powder in PBST added for blocking. The plates were incubated for ~ h at temperature on a shaker. The plates were washed with PBST, ul / well solutions from 2nd and 3rd rounds of selection with at 1:2 to 1:256 was added to the plates. were for © h at room temperature on a shaker. were coated with anti-M13 the plates were washed, 50 nl / well of reaction and TMB) in acetate buffer pH 5.0 was added. were at room temperature in dark until a slight appeared the negative controls (but for not more than 20 The reaction was quenched by adding 25 ul / well of 10% H2S04, OD wells was measured at 450 nm. The assay revealed specific (5X the background of © human naive Fab library and 6 immune Fab CHARM 3rd round of selection on target antigen and absence of (less than 2X the background signal) non-specific binding. The antibody variable domains genes from these libraries recloned into expression vector pLL-Fab (Fig. 8) to produce secretable soluble form of Fab in E. coli cells. Example 6 Screening of Fabs that specifically bind human GITR ELISA was used to detect Fabs that specifically bind human which are secreted into the medium from monoclones produced in E. according to Example 5. Fab with a published sequence, (patent application), was used as a positive control. For binding assay, ELISA well plates (medium binding, Greiner were coated with 50 pl / well of GITR (0.2 pg / ml in 1X carbonate sealed and incubated overnight at 4°C. All further stages performed in accordance with standard ELISA protocols using a performance automated platform based on robotic systems such as Qpix2xt (Molecular Devices) and Tecan Freedom EVO 200 (Tecan) specific binding was blocked by adding a blocking buffer BB (200 0.5% fat-free milk in PBS). Plates were incubated on a shaker for at room temperature. After washing with PBS-Tween, each cell was with 50 pl of test Fab-containing cell supernatant mixed with the volume of BB. Plates were incubated on a shaker for I hour temperature; further, each plate well was 3 times Tween buffer. After washing, each well was coated anti-human Fab HRP-conjugated secondary ThermoScientific) in PBS-Tween (1:5000)}. Plates were on rotation shaker (50 min at room temperature) and then 3 with PBS-Tween buffer as described above. Colorimetric developed by adding TMB (50 pl / well) until saturation (average min); further color development was blocked by adding a stop (30 pl / well, 10% sulfuric acid). Color signal was measured at using an appropriate Tecan-Sunrise plate reader (Tecan). binding level was proportional to color signal production. Clones, which the color signal 5X exceeded the background signal and comparable to that of the control antibody Fab-gitr3215, were by ELISA to detect non-specific binding. Example 7 Non-specific binding assay of selected Fab-other interactions ELISA was also employed to assay non-specific binding of fragments in question to other antigens. The study was performed described but IL6R-Fc, INFo2b, PCSK9-VG-FE, PD-1-Fc (2.5 in 1x buffer) were used as immobilization antigens. TEV-Fc (0.2 in 1x carbonate buffer) was as & the standard ELISA protocol using a high-performance platform based on robotic systems such as Genetix Qpix2xt Devices) and Tecan Freedom EVO 200 (Tecan). Clones, in which signal of nonspecific binding did not exceed the had 5X lower values as compared to those of considered positive and their genes were sequenced determination of antibody variable domain gene determination of uniqueness. As a result of sequencing, we 30 unique sequences for conversion to a full-length IgGl format. Example 8 Production of recombinant antibodies in suspension culture mammalian cells Antibody variable domain genes from Example 7 were according to standard methods. To this end, we generated PCR containing antibody heavy and light chain variable domain genes. heavy chain variable domain was cloned into vector pEE-Hc IgGl SalI / Nhel restriction sites, which schematic map is shown in Fig. The light chain variable domain was ligated into vector pEE-CK SalIl / BsiWI restriction sites, which schematic map is shown in Fig. Antibodies were generated in established cell line obtained Chinese hamster ovary cells (CHO-K1) according to published [Biotechnol Bioeng. 2005 Sep 20; 91(6):670-677, Liao Biotechnol Lett. 2006 Jun;28(11):843-848; Biotechnol Bioeng. 5;84 (3) :332-342]. Cells constitutively expressing the EBNAL Barrvirus nuclear antigen 1) protein gene were used. The CHO-K1-S cell line cells were used to generate antibodies a transient expression system. Cells were cultured in baffled (125, 250, 500, 1000 and 3000 ml) in a mixture II FreeStyle CHO (1:1) media supplemented with 4 mM gentamicin and 10 ug / ml ciprofloxacin in at +37C, 5% C02, and 110 or 150 on a were subcultured 3 times a week, 0.2*106 For transfection, cells were the day transfection at a density of 0.8*106 transfection performed one day later using a cell culture at a density of cells / ml. RPMI-1640 medium supplemented with 2 mM glutamine mg / ml of gentamicin was used to prepare the transfection Vectors were diluted separately in the medium at 0.75 transfection agent, polyethyleneimine (PEI), was diluted at PEI:DNA 7:1 by weight. The dilutions of vectors and PEI were and incubated for 10 minutes at room temperature, the mixture was then introduced to the cells, and the cells were under standard conditions. The next day after transfection, a mixture of CHO-S-SFM FreeStyle CHO (1:1) media supplemented with 0.05 mg / ml of and 10 ng / ml of ciprofloxacin, ~. mM sodium valproate and F12.7 was added to the cells, the cells were shaker-incubators at +34C, 5%C0O2 and 110 or 150 rpm on flask size. On day 3-4 after transfection, 10% feeding was to the On after cell samples were (Table 2). Recombinant antibodies were isolated and purified culture liquid using Protein A affine HPLC column. Cleared liquid was passed through ~ ml HiTrap rProtein A FF column Healthcare) that was equilibrated with phosphate buffered saline pH 7.4). Then the column was washed with 5 column volumes remove any non-specifically binding components. Bound eluted using 0.1 M glycine buffer (pH 3). The principal peak was collected and brought to neutral pH with 1 M 8). All stages were conducted under 110 cm / h flow rate. The was then dialyzed into PBS (pH 7.4) by means of SnakeSkin Tubing technique, filtered (0.22 um), transferred into tubes at -70°C. Purity the resulting protein solution was evaluated by SDS (Figs. 11 and 12). All antibodies comply with level for examining physico-chemical properties and in Example 9 Kinetic assay of anti-GITR IgGl antibody-human GITR Binding affinity constants for anti-GITR macaque / cynomolgus monkey GITR were OctetRed 96 according to ug / ml was non-specifically onto surface reactive sensors of the generation standard protocol to manufacturer’s preparation and of AR2G sensors). of antibodies The were added at a concentration The assay conducted at PBS comprising 0.1% Tween-20 and 0.1% BSA a working buffer. The resulting sensograms, after subtracting a were analyzed using Octet Data Analysis software accordance with the standard procedure and using 1:1 The resulting affinity constants are shown in Table 3. All antibodies show high affinity and specificity for human GITR. 3. Affinity constants for anti-GITR antibody - human Example 10 Determination of anti-GITR specific agonist activity. For the assay, we used HEK293-GITR-NFkB-Luc cell line on the basis of Hek-293 cell line, stably expressing on surface and containing the firefly luciferase encoding gene control of NFkB promoter. The assay was conducted in a 96-well culture plate. The in each well contained HEK293-GITR-NFkB-Luc cells and the test at a concentration as indicated in the graph. All suspension were prepared in a nutrient cell culture medium. After components, the plate was incubated at 37°C, 5%CO: and luminescence assay kit and a plate reader, the luciferase in the wells was measured. Examination of specific activity of anti-GITR antibodies revealed (Fig. 13) that the antibodies of lots and 1213 are functionally active and exhibit significant activity with an upper plateau comparable to that of both the antibody anti-GITR3215 antibody and GITRL. These antibodies renamed according to Table 4. Example 11 Enzyme-linked immunoassay of interactions between antibodies and GITR from different organisms ELISA was used to measure relative affinity of antibodies GITR-Fc from different organisms. For binding assay, ELISA plate (medium binding from Greiner bio one) were coated with 50 human / cynomolgus / murine GITR-Fc (0.5 pg / ml in 1x carbonate sealed and incubated overnight at 4°C. All further steps in accordance with the standard ELISA protocol described 6. Anti-GITR antibody BCD166-01-01 specifically bound to the human cynomolgus monkey GITRs and did not bind to the murine GITR (Fig. Antibody BCD166-01-011 specifically bound to the human GITR, but not exhibit binding to the cynomolgus monkey GITRs 15). Antibody BCD166-01-014 specifically bound to binding to the cynomolgus monkey GITR was much weaker, and no binding was observed for the murine GITR (Fig. 16). candidate BCD166-01-011 was excluded from further study due to lack of apparent specificity for the monkey GITR. Candidates 01-01 and BCD166-01-014 were finalized for further development. Example 12 Kinetic assay of anti-GITR IgGl antibody macaca GITR / macaca Cynomolgus GITR interactions Binding affinity constants for anti-GITR antibodies and macaque / cynomolgus monkey GITR were obtained using OctetRed according to manufacturer's (ForteBio) protocol. Antigen 25 ng / ml non-specifically immobilized onto the surface of sensors of the second generation (AR2G) (ForteBio, protocol according to manufacturer’s instructions for immobilization of AR2G sensors). of anti-GITR antibodies were added at a concentration The assay was 30°C using PBS comprising 0.1% Tween-20 and 0.1% BSA as a buffer. The resulting sensograms, after subtracting a were analyzed using Octet Data Analysis software accordance with the standard procedure and using 1:1 The resulting affinity constants are shown in Tables 6. tested antibodies exhibit high affinity and specificity for GITR, thus providing the basis for further research on this in vivo animal model. Table 5. Affinity constants of anti-GITR antibody - macaca mulatta Table 6. Affinity constants of anti-GITR antibody - macaca Cynomolgus Example 13 In vitro cellular assay for determination of anti-GITR agonist activity of wild and mutant candidates with enhanced activity. It has a can to of on surfaces after binding to and enhance functions, such as ADCC, The aim was to obtain an but not CDC. Mutagenesis was to engineering protocols, the was according to Example 8. The was is hereinafter as BCD166-02-01. Two parent BCD166-01-014 were also assayed. The assay was performed in the same fashion as that of 10. The results shown in Fig. 17 indicate a 5-fold increase in level of activation in cell agonist assay of test BCD166-02-01 compared to precursor BCD166-01-01, to which the E345R mutation introduced, which arises from antigen-dependent oligomerization anti-GITR antibody BCD166-02-01 on the target cell. Moreover, not the level of the upper plateau of activation naturally increased, also the EC50 value increased more than 20 times in the direction gain. Example 14 In vitro cellular assay for antibody-dependent mediated cytotoxicity (ADCC) of anti-GITR antibodies. In the assay, we studied ADCC activity for candidates 01. Two antibodies BCD166-01-01, BCD166-01-014 were also Furthermore, by analogy to candidate BCD166-01-01, the E345R was introduced into candidate BCD166-01-014 (which was called 02-014) to increase ADCC activity relative to that of the wild In the assay, we used Jurkat-NFAT-Luc-CD16 cell line on Jurkat cell line, stably expressing CD16 on the surface containing the firefly luciferase encoding gene under control of promoter; and Hek-293-GITR cell line created based on Hek-293 line, stably expressing GITR on the surface. The assay was conducted in a 96-well culture plate. The in each well contained Jurkat-NFAT-Luc-CD16 effector cells 293-GITR target «cells, as well as the test antibodies at concentration as indicated in the graph. All suspension were prepared in a nutrient cell culture medium. After adding components, the plates were incubated at 37°C, 5%CO. and a luciferase assay kit and a plate reader, we measured the intensity in the wells. The results shown in Fig. 18 indicate a 40-fold in EC50 value in ADCC assay of test BCD166-02-01 as that to which the was antibody BCD166-02-01 on the target cell. A 5-fold increase in EC50 value was also reliably detected in ADCC of 02-14 as compared to that of precursor BCD166-01-14, mutation was introduced. The aggregate data clearly significant positive effect of the E345R substitution in the of IgGl antibodies on different effector characteristics necessary the production of a highly active GITR agonist. Example 15 In vitro cellular assay for compement-dependent (CDC) of anti-GITR antibodies. In the assay, we studied antibodies BCD166-01-01, BCD166-02-01 and control anti-GITR-3215 antibody. The assay was conducted in a 96-well culture plate. The in each well comprised HEK-293-GITR cells, as well as test at an indicated concentration and human serum complement. containing the described suspension was incubated for 4 h at CO. Further, alamar blue solution was added to each well, the was then incubated at 37°C, 5% CO2. Further, using a plate fluorescence intensity was measured in wells (excitation of 544 nm, emission wavelength of 590 nm). The results shown in Fig. 19 indicate CDC i.e. not more than 10% to a concentration which limit concentration for estimating a therapeutic use in humans, for antibodies BCD166-01-01, anti-GITR-3215 antibody. For candidate BCD166-01-14, a concentration of © pg / ml was about 30% lysis, thus apparent effect, but possibly modest in vivo. The aggregate clearly indicate an insignificant effect of the E345R substitution the Fc portion of anti-GITR IgGl antibody candidates. Example 16 In vitro cellular assay of anti-GITR The assay was to evaluate the negative or effect of test candidates on the change in different responder cells. In we antibodies BCD166-01-014, CD20 antibody PBMCs were density gradient assay was a culture plate. The each well PBMCs as indicated in the graph at a of 25, * and 0 After mixing PBMCs and antibodies, was incubated for at 37°C, 5% C02. Then, the proportion CD19+, CD4+ CD8+ subpopulations of PBMCs in was staining of the suspensions against the corresponding CDs and subsequent cell flow cytofluorometer. For CD56+, CD19%+, CD3+ cells, the proportion thereof relative to all cells of test whereas for CD4+, CD8+, the graphs show the proportion to CD3+ cells. The results shown in Figs. 20-24 indicate a negative effect of test anti-GITR candidates, i.e. not more reduction in number of responder cells as compared to the In the case of NK cell we observed about antibody, however, also showed an increase in NK cell virtually natural complete depletion of CD20+ B cell all candidates in question do not exhibit cytotoxicity toward human blood cells. Example 17 In vitro cellular assay for antibody-dependent depletion of nTreg cell population. ADCC-dependent depletion of GITR* nTreg cell population contemplated to be one of the main contemplated mechanisms for therapeutic anti-oncogenic effect of anti-GITR antibodies. In assay, we studied antibodies BCD166-01-01, BCD166-01-014, 01 and control anti-CTLA4 antibody (ipilimumab) . Peripheral blood mononuclear cells (PBMCs) were whole blood from healthy donors by Ficoll centrifugation. PBMCs were enriched for nTregs Regulatory T Cell Isolation Kit, human” (Miltenyi The resulting cells were activated using immobilized anti-CD3 and anti-CD28 antibodies. NK cells were isolated from PBMCs using "NK Cell Isolation human" (Miltenyi Biotec, Germany). Antibody solutions, NK cell suspension and nTreg suspension added to a culture plate. The plate was incubated at 37 °C, 5% CO: 16 hours. Samples of cell suspensions were stained using labeled antibodies against CD3, CD4, CD25, FoxP3 (Biolegend, Samples of stained «cell suspensions were analyzed by cytofluorometry. The results of analysis for antibody-dependent under effect of test antibodies shown in Fig. 25 for cellular material and in Fig. 26 for donor 2 indicate 70-100% decrease in cell population, i.e. depleting of anti-GITR candidates, especially for the donor 2 Furthermore, a comparative analysis with the control antibody ipilimumab shows that anti-GITR antibodies activity, especially the final candidate BCD166-02-01. Example 18 In vitro cellular assay for antibody-dependent depletion of iTreg cell population. ADCC-dependent depletion of GITR+ iTreg cell population contemplated to be one of the main contemplated mechanisms for therapeutic anti-oncogenic effect of anti-GITR antibodies. In assay, we studied antibodies BCD166-01-01, BCD166-01-014, 01. PBMCs were isolated from whole blood from healthy donors density gradient centrifugation. Monocytes as a population plastic-binding cells were isolated from PBMCs. To obtain cells, monocytes were incubated in the presence of 1000 GM-CSF (Peprotech, USA) and 500 units / ml of IL-4 (Thermo USA) at 37°C 5% CO» for 120 hours. LPS solution (Sigma, USA) was to the culture medium to a concentration of 0.5 ug / ml, the cells incubated at 37°C 5% CO, for 48 hours. Antibody dilution solutions, dendritic cell suspension were added to the culture plate. The plate was incubated at 37 CO, for 120 hours. Samples of cell suspensions were stained using labeled antibodies against CD3, CD4, CD25, FoxP3 (Biolegend, Samples of stained cell suspensions were analyzed by cytofluorometry. Anti-GITR antibodies BCD166-01-01, BCD166-01-14 BCD166-02-01 induce iTreqg depletion in vitro. The results of assay for antibody-dependent iTreg effect of test antibodies shown in Fig. 27 for the cellular material indicate a significant 2-5-fold of test anti-GITR candidates, especially for the donor 1 final candidate BCD166-02-01 shows sufficient iTreg activity. Example 19 In vitro cellular assay for secretion cytokines IL-2 and IFN-vy under the effect of human cell population. PBMCs were isolated density gradient centrifugation. Anti-CD3 and anti-GITR antibodies were immobilized in the of a 96-well plate. To this end, 100 ul / well of DPBS containing anti-GITR antibody BCD166-02-01 at a indicated in the graph and anti-CD3 antibody at concentration were added to corresponding plate wells, the incubated for 16 hours at room temperature. The assay was performed using a 96-well plate with anti-GITR antibodies pre-immobilized in the wells. The each well contained 40,000 PBMCs and anti-CD28 antibody at concentration.All suspension components were prepared in medium containing 10% FBS. After adding the cell was incubated for 6 days at 37°C, 5% C02. On aliquots of culture liquid were collected from wells. Then, concentrations of IL-2 and IFN-y were measured culture liquid day 4 and 6 of incubation using ELISA. Analysis of the effect of anti-GITR antibodies on secretion of proinflammatory cytokines IL-2 and IFN-y anti-cancer effect showed a significant increase in the of these substances in culture liquid (the 28). Thus, candidate BCD166-02-01 ‘exhibits secretion activation, which may indicate a effect. 20 for BCD166-02-01~- constants for BCD166-02-01- were receptors were on sensors (SA). We conducted and analyzed the dissociation between receptors and antibody in a buffer containing 0.1% Tween-20 and 0.1% BSA: for FcRn receptor we buffer, for the other receptors we used pH 7.4 buffer) resulting sensograms were analyzed in accordance with or models, the affinity constants were calculated using Octet Analysis Software 8.0 User Guide Copyright 20110. "Results of affinity assay for BCD166-02-01- FcRn,FcgRIIIal58V,FcgRIIIal58F, FcgRIIal3lH, FcgRIIal3lR, FcgRIIb FcgRIa interactions are shown in Fig. 23. They amplified values compared to the literature repeatedly measured in our company for IgGl isotype indicating a noticeable but not qualitative effect of mutation in the Fc fragment on the kinetics of monomeric adsorbed on these receptors from solution. Example 21 Determination of colloidal and thermal stability by aggregation point using dynamic light scattering In order to determine the aggregation temperature of under study by dynamic light scattering, dependence of in the medium on temperature was obtained using heating 40 to 85 Table 7. BCD166-02-01 aggregation point One may conclude that molecule BCD166-02-01 has a high colloid stability (aggregation point in 20mM Acetate, pH = 20mM His, pH = 5.5 buffer solutions is > 65°). Similar data were obtained for BCD166-01-014. Example 22 Determination of thermal stability under 50°C thermal stress The test were placed in a thermostated air bath thermostated at 72 hours. After heating, intact samples were size-exclusion HPLC (SEC a detector and by isoelectric method. was on 1100 HPLC system on column, was performed at nm. was determined by Labchip GX II, chip was carried to Charge Variant and Kit, PerkinElmer 4 The resulting data for BCD-166 stability (for BCD166-01-014) under 50°C incubation are shown in Table 8; (for BCD166-02-01) and 31 (for BCD1l66-01-014) show chromatograms under 72-hour incubation at 50°C. conclusion: the sample has high colloidal and * A is the difference between purity of a stressed sample purity of an intact sample (input control), %. Table 8. Dependence of monomer content by size-exclusion HPLC electrophoresis for BCD-166. Example 23 Assessment of anti-tumor activity of BCD-166 products using subcutaneous xenograft model Antitumor activity of BCD-166 products subcutaneous tumor xenograft model. To this line cells in admixture with Matrigel® were humanized huNOG-EXL mice. On day 7 after cell were divided in groups as shown in Table 9. Table 9. Groups of animals in the study of anti-tumor on day 1, 5, 8, 12 after buffer was adminitered to group. and tumor linear dimensions were measured throughout the Tumor volume was calculated using the formula V=LxWxHxm / 6. of test product was assessed by the index of tumor growth (TGI) that was calculated taking into account the mean tumor in the negative control group (Vc) and the group of product (Vt) the fc The experiment revealed high anti-tumor activity of 014 and BCD-166-02-01 products, whereas BCD-166-01-01 product show any significant anti-tumor activity. The results are Figs. 32 and 33.

Claims

Claims:

1. An isolated monoclonal antibody or antigen-binding fragment thereof specifically binds to GITR comprising: (a) a heavy chain variable domain comprising: (1) CDR1 comprising an amino acid sequence selected from the group: NYGMH (SEQ ID NO: 1) or YYWMY (SEQ ID NO: 12); (ii) CDR2 comprising an amino acid sequence selected from the group: VIWFDGSNKFYTDSVKG (SEQ ID NO: 2) or AISWNGGRTYYAESMKG (SEQ ID NO: (iii) CDR3 comprising an amino acid sequence selected from the ELGGYYYDSSGFRPYYYGMDV (SEQ ID NO: 3) or NRYYSDPNYGMNL (SEQ ID NO: and (b) a light chain variable domain comprising: (1) CDR1 comprising an amino acid sequence selected from the group: RASQSIGSWLA (SEQ ID NO: 7) or TGTSTDIGTYKYIS (SEQ ID NO: 17); (ii) CDR2 comprising an amino acid sequence selected from the group: AASTLQOR (SEQ ID NO: 8) or GVSHRPS (SEQ ID NO: 18); (iii) CDR3 comprising an amino acid sequence selected from the QQSHSHPLT (SEQ ID NO: 9) or SSYTSSGTVV (SEQ ID NO: 19).

2. A monoclonal antibody or antigen-binding fragment thereof according claim 1, wherein the heavy chain variable domain comprises: (1) CDR 1, CDR 2 and CDR 3 acid sequences represented by the of SEQ ID NO: 1, SEQ ID NO:2 and SEQ ID NO:3, respectively. (ii) CDR 1, CDR 2 and CDR 3 acid sequences represented by the of SEQ ID NO: 12, SEQ ID NO:13 and SEQ ID NO:14, respectively.

3. A monoclonal antibody or antigen-binding fragment thereof according claim 1, wherein the light chain variable domain comprises: (1) CDR 1, CDR 2 and CDR 3 comprising amino acid sequences by the sequences of SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9, (ii) CDR 1, CDR 2 and CDR 3 comprising amino acid sequences by the sequences of SEQ ID NO:17, SEQ ID NO: 18 and SEQ respectively.

4. A monoclonal antibody or antigen-binding fragment thereof according claim 1, wherein the heavy chain variable domain comprises CDR 1, CDR 2 and CDR comprising amino acid sequences represented by the sequences of SEQ ID 1, SEQ ID NO:2 and SEQ ID NO: 3, respectively; the light chain variable domain comprises CDR 1, CDR 2 and CDR comprising amino acid sequences represented by the sequences of SEQ ID 7, SEQ ID NO:8 and SEQ ID NO:9, respectively.

5. A monoclonal antibody or antigen-binding fragment thereof according claim 1, wherein the heavy chain variable domain comprises CDR 1, CDR 2 and CDR comprising amino acid sequences represented by the sequences of SEQ ID 12, SEQ ID NO: 13 and SEQ ID NO: 14, respectively; the light chain variable domain comprises CDR 1,CDR 2 and CDR comprising amino acid sequences represented by the sequences of SEQ ID 17, SEQ ID NO: 18 and SEQ ID NO: 19, respectively.

6. A monoclonal antibody or antigen-binding fragment thereof according claim 1, wherein the heavy chain variable domain comprises: (1) an amino acid sequence that is at least 90% homologous to the acid sequence of SEQ ID NO: 4; or (ii) an amino acid sequence that is at least 90% homologous to amino acid sequence of SEQ ID NO:

15.

7. A monoclonal antibody or antigen-binding fragment thereof according claim 1, wherein (1) the heavy chain variable domain comprises the amino acid of SEQ ID NO:

4. (ii) the heavy chain variable domain comprises the amino acid of SEQ ID NO:

15.

8. A monoclonal antibody or antigen-binding fragment thereof according claim 1, wherein the light chain variable domain comprises: (1) an amino acid sequence that is at least 90% homologous to the acid sequence of SEQ ID NO: 10; or (ii) an amino acid sequence that is at least 90% homologous to amino acid sequence of SEQ ID NO: 2C.

9. A monoclonal antibody or antigen-binding fragment thereof according claim 1, wherein (1) the light chain variable domain comprises the amino acid of SEQ ID NO: 1C; or (ii) the light chain variable domain comprises the amino acid of SEQ ID NO: 2C.

10. A monoclonal antibody or antigen-binding fragment thereof according claim 1, wherein (i) - the heavy chain variable domain comprises an amino acid sequence is at least 90% homologous to the amino acid sequence of SEQ ID NO: 4; - the light chain variable domain comprises an amino acid sequence is at least 90% homologous to the amino acid sequence of SEQ ID NO: 10; (ii) - the heavy chain variable domain comprises an amino acid sequence is at least 90% homologous to the amino acid sequence of SEQ ID NO: 15; - the light chain variable domain comprises an amino acid sequence is at least 90% homologous to the amino acid sequence of SEQ ID NO:

20.

11. A monoclonal antibody or antigen-binding fragment thereof according claim 1C, wherein - the heavy chain variable domain comprises the amino acid sequence SEQ ID NO: 4; - the light chain variable domain comprises the amino acid sequence SEQ ID NO: 1C.

12. A monoclonal antibody or antigen-binding fragment thereof according claim 1C, wherein - the heavy chain variable domain comprises the amino acid sequence SEQ ID NO: 15; - the light chain variable domain comprises the amino acid sequence SEQ ID NO:

20.

13. A monoclonal antibody according to claim 1, wherein the antibody specifically binds to GITR is a full-length IgG antibody.

14. A monoclonal antibody according to claim 13, wherein the full-length antibody relates to human IgGl, IgG2, IgG3, IgG4 isotype.

15. A monoclonal antibody according to claim 14, wherein the full-length antibody relates to human IgGl isotype.

16. A monoclonal antibody according to claim 1, wherein the specifically binds to GITR comprises the E345R mutation in the Fc to increase agonist properties, antibody-dependent cellular (ADCC), but not complement-dependent cytotoxicity (CDC).

17. A monoclonal antibody according to claim : comprising a heavy comprising: (1) an amino acid sequence that is at least 90% homologous to sequence of SEQ ID NO: 5; or (ii) an amino acid sequence that is at least 90% homologous to sequence of SEQ ID NO:

6.

18. A monoclonal antibody according to claim 1 comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: or (ii) a heavy chain comprising the amino acid sequence of SEQ ID NO:

19. A monoclonal antibody according to Claim : comprising a light comprising an amino acid sequence that is at least 90% homologous to sequence of SEQ ID NO:

11.

20. A monoclonal antibody according to Claim [ comprising a light comprising the amino acid sequence of SEQ ID NO:

11.

21. A monoclonal antibody according to Claim 1 comprising: ti) - a heavy chain comprising an amino acid sequence that is at least homologous to the sequence of SEQ ID NO: 5; - a light chain comprising an amino acid sequence that is at least homologous to the sequence of SEQ ID NO: 11; or (ii) - a heavy chain comprising an amino acid sequence that is at least homologous to the sequence of SEQ ID NO: 6; - a light chain comprising an amino acid sequence that is at least homologous to the sequence of SEQ ID NO:

11.

22. A monoclonal antibody according to claim 1 comprising: - a heavy chain comprising the amino acid sequence of SEQ ID NO: 5; - a light chain comprising the amino acid sequence of SEQ ID NO:

11.

23. A monoclonal antibody according to claim 1 comprising: - a heavy chain comprising the amino acid sequence of SEQ ID NO: 6; 24. A monoclonal antibody according to claim [ comprising a heavy comprising an amino acid sequence that is at least 90% homologous to sequence of SEQ ID NO:

16.

25. A monoclonal antibody according to claim I comprising a heavy comprising the amino acid sequence of SEQ ID NO:

16.

26. A monoclonal antibody according to claim [ comprising a light comprising an amino acid sequence that is at least 90% homologous to sequence of SEQ ID NO:

21.

27. A monoclonal antibody according to claim [ comprising a light comprising the amino acid sequence of SEQ ID NO:

21.

28. A monoclonal antibody according to claim 1 comprising: - a heavy chain comprising an amino acid sequence that is at least homologous to the sequence of SEQ ID NO: 16; - a light chain comprising an amino acid sequence that is at least homologous to the sequence of SEQ ID NO:

21.

29. A monoclonal antibody according to claim 1 comprising: - a heavy chain comprising the amino acid sequence of SEQ ID NO: 16; - a light chain comprising the amino acid sequence of SEQ ID NO:

21.

30. An isolated nucleic acid encoding an antibody or antigen-binding thereof according to any of claims 1-29.

31. A nucleic acid according to claim 3C, wherein the nucleic acid is 32. An expression vector comprising a nucleic acid according to 30-31.

33. A method of obtaining a host cell binding fragment thereof according to any claims transformation of a cell with a vector according to claim 32.

34. A host cell for obtaining of an antibody or antigen-binding thereof according to any of claims 1-29 comprising a nucleic acid to any of claims 30-31.

35. A method of obtaining of an antibody or antigen-binding fragment according to any of claims 1-29 comprising culturing of a host cell to claim 34 in a culture medium under conditions sufficient to obtain antibody, if necessary, followed by isolation and purification of obtained antibody.

36. A pharmaceutical composition for treatment of a disease or mediated by GITR comprising an antibody or antigen-binding fragment according to any of claims 1-29 in a therapeutically effective amount combination with one or more pharmaceutically acceptable excipients.

37. A pharmaceutical composition according to claim 36 of a disease or disorder mediated by GITR selected from the group: cancer, head and neck cancer, stomach cancer, breast cancer, renal cancer, CRC (colorectal cancer), (OC) ovarian cancer, NSCLC (non-small lung cancer).

38. A pharmaceutical composition for treatment of a disease or mediated by GITR comprising an antibody or antigen-binding fragment according to any of claims 1-29 in a therapeutically effective amount least one therapeutically active antitumour compound in a effective amount.

39. A pharmaceutical composition according to claim 38 of a disease or disorder mediated by GITR selected from the group cancer, head and neck cancer, stomach cancer, breast cancer, renal cancer, CRC (colorectal cancer), (OC) ovarian cancer, NSCLC (non-small lung cancer).

40. A pharmaceutical composition according to claim 38, wherein therapeutically active antitumour compound is selected from chemotherapeutic agent, antibody or anti-hormonal agent.

41. A pharmaceutical composition according to claim 40, wherein therapeutically active antitumour compound is (i) an antibody selected from the group: anti-PDl1 antibodies, L1 antibodies, anti-CTLA4 antibodies, anti-4-1BB antibodies, antibodies or combinations thereof, or (ii) a small molecule, or (iii) selected from the group of activators of innate or immunity.

42. A pharmaceutical composition according to claim 41, wherein the according to claim 29 and the at least one therapeutically active compound are administered sequentially.

43. A pharmaceutical composition according to claim 41, wherein the according to claims 1-29 and the at least one therapeutically antitumour compound are administered simultaneously.

44. A method for inhibition of biological activity of GITR in a subject need of such inhibition comprising administration of an effective amount an antibody or antigen-binding fragment thereof according to any of 1-29.

45. A method for treatment of a disease or disorder mediated by comprising administration in a subject in need of such treatment of antibody or antigen-binding fragment thereof according to any of claims 29 or a pharmaceutical composition according to claims 36 or 38 a therapeutically effective amount.

46. A method for treatment of a disease or disorder according to claim wherein the disease or disorder is selected from the group: cervical head and neck cancer, stomach cancer, breast cancer, renal cell cancer, (colorectal cancer), (OC) ovarian cancer, NSCLC (non-small cell 47. Use of an antibody or antigen-binding fragment thereof according to of claims 1-29 or a pharmaceutical composition according to claims 36 or for treatment in a subject in need of such treatment of a disease mediated by GITR.

48. Use according to claim 47, wherein the disease or disorder is from the group comprising: cervical cancer, head and neck cancer, cancer, breast cancer, renal cell cancer, CRC (colorectal cancer), ovarian cancer, NSCLC (non-small cell lung cancer).

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