Multispecific deimmunising cd3-binders

FIELD: chemistry; biochemistry.

SUBSTANCE: present invention relates to immunology and biotechnology. The invention discloses versions of a cytotoxically active CD3-specific binding structure. The structure comprises a first domain specifically binding to human CD3 and an Ig-derived second binding domain which is specific to molecules on the cell surface. The invention describes a coding nucleic acid, a vector for expressing the structure and an eukaryotic cell transformed by the vector. The invention discloses versions of compositions based on the structure for treating, preventing or alleviating various diseases and corresponding methods of treating the diseases. A method of obtaining the structure is disclosed.

EFFECT: use of the invention provides a structure with low immunological potency, which has cytotoxicity comparable to the initial structure, which may find further use in medicine.

60 cl, 18 dwg, 15 tbl, 8 ex

 

The text descriptions are given in facsimile form.

1. CD3 specific binding construct, with cytotoxic activity, including the first domain specifically binding to human CD3, and the second binding domain derived from Ig, in which said first domain is neimmunizirovannah and includes the region CDR-H1 containing the amino acid sequence shown in the sequence SEQ ID NO:88, region CDR-H2 containing the amino acid sequence shown as sequence SEQ ID NO:90 and SEQ ID NO.:93 and region CDR-H3, containing the amino acid sequence shown as sequence SEQ IDNO.:96, 108, 119, 120, 121, 122, 123, 124, 125, 126 or 127; and in which said first domain also includes in its frame H1 sequence reserve a ticket, and also a transitive sequence between the frame H1 and area CDRH1 comprises the sequence Ala-Ser-Gly-Tyr-Thr-Phe (ASGYTF; SEQ ID NO.:233), a specified second domain derived from Ig, includes the site of interaction with a molecule on the surface of cells with specificity against the specified molecule.

2. CD3 specific binding construct, with cytotoxic activity according to claim 1, additionally including the specified first domain frame H3 comprising the sequence Met-Glu-Leu-Ser (MELS; SEQ ID NO.:234).

3. CD3 specific binding construct, with cytotoxic activity, according to claim 1, additionally including the specified first domain frame H3 comprising the sequence Ile-Thr-Thr-Asp-Lys (ITTDK; SEQ ID NO.:235).

4. CD3 specific binding construct according to claim 1, in which said first domain specifically binding to human CD3, includes a frame H1, shown in the sequences SEQ ID NO.:152 or 153.

5. CD3 specific binding construct according to claim 1, in which said first domain specifically binding to human CD3, includes a frame H2, shown in the form is of posledovatelnostei SEQ ID NO.:156 or 157.

6. CD3 specific binding construct according to claim 1, in which said first domain specifically binding to human CD3, includes a frame H3, shown in the sequences SEQ ID NO.:160 or 161.

7. CD3 specific binding construct according to claim 1, in which said first domain specifically binding to human CD3, includes a frame H4, shown in the sequences SEQ ID NO.: 164 or 165.

8. CD3 specific binding construct according to claim 1, in which the design includes the region VHshown in the sequences SEQ ID NO.:74 or 76.

9. CD3 specific binding construct according to claim 1, in which the said construction comprises CDR-L1, shown in the sequences SEQ ID NO.:98 or 100.

10. CD3 specific binding construct according to claim 1, in which the design includes CDR-L2, shown in the form of sequence SEQ ID NO.:102.

11. CD3 specific binding construct according to claim 1, in which the design includes CDR-L3, as shown in the sequence SEQ ID NO.:104.

12. CD3 specific binding construct according to claim 1, including the region VLin my CD3-specific parts, in which the specified region VLselected from the group comprising sequence SEQ ID NO.:78, SEQ ID NO.:80, SEQ ID NO.:82 and SEQ ID NO.:112.

13. CD3-specific binding design by p., in which specified a second domain derived from Ig, is a scFv.

14. CD3 specific binding construct according to claim 1, in which the specified second domain derived from Ig, and/or (a) binding of the linker area (s) is (are) humanitarianism and/or deimmunized.

15. CD3 specific binding construct according to 14, in which the molecule on the surface of the cell is a tumor-specific marker.

16. CD3 specific binding construct according to 14, in which the specified second binding domain derived from Ig, includes the site of interaction with the antigen having specificity against molecules selected from the group consisting of Arcam, CCR5, CD19, HER-2, HER-2 neu, HER-3, HER-4, EGFR, PSMA, CEA, MUC-1 (mucin), MUC2, MUC3, MUC4, MUC5AC, MUC5B, MUC7, antigen, Lewis-Y, CD20, CD33, CD30, ganglioside GD3, 9-O-acetyl-GD3, GM2, Globo H, fucosyl GM1, poly SA, GD2, carbonic anhydrase IX (MN/CA IX), CD44v6, Sonic Hedgehog (Shh), Wue-1, plasmacytomas antigen (membrane-bound) IgE, chondroitin sulfate of melanoma proteoglycan (MCSP), CCR8, the precursor of TNF-alpha, STEAP, mesothelin, A33 antigen, stem cell antigen of the prostate (PSCA), Ly-6;
desmoglein 4, a neo-epitope of E-Katerina, receptor fetal acetylcholine, CD25, a marker CA19-9 marker CA-125 and receptor type II Muellerian-inhibiting substance (MIS), sTn (sieciowego antigen, TAG72), FAP (antigen activation fibres which blasts), indocyanine, EGFRvIII, L6, SAS, CD63, TAG72, TF-antigen measles antigen CD7, CD22, Igα, Igβ, G250, gp100, MT-MMPs, F19-antigen, CO-29 and EphA2.

17. CD3 specific binding construct according to claim 1, in which the specified second binding domain derived from Ig, includes the site of interaction with the antigen having specificity against Arcam.

18. CD3 specific binding construct according to 17, in which the CD3 specific binding construct comprises the amino acid sequence selected from the group
(a) the amino acid sequence represented by one of the sequences SEQ ID NO.:31, 33, 35, 37, 39, 49, 55, 58, 61, 63, 65, 67, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323 and 325;
(b) amino acid sequence encoded by the nucleic acid sequence represented by one of the sequences SEQ ID NO.:30, 32, 34, 36, 38, 48, 54, 57, 60, 62, 64, 66, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322 and 324; and
(C) the amino acid sequence encoded by a nucleic acid sequence which is degenerate as a result of the genetic code to the nucleotide sequence of (b).
(g) the amino acid sequence encoded by the sequence NUS is einevoll acid, hybridises with complementary chain nucleic acid sequence described in (b) under conditions of stringent hybridization and having specificity against CD3 and molecules selected from the group consisting of Arcam, CCR5, CD19, HER-2, HER-2 neu, HER-3, HER-4, EGFR, PSMA, CEA, MUC-1 (mucin), MUC2, MUC3, MUC4, MUC5AC, MUC5B, MUC7, antigen, Lewis-Y, CD20, CD33, CD30, ganglioside GD3, 9-O-acetyl-GD3, GM2, Globo H, fucosyl GM1, poly SA, GD2, carbonic anhydrase IX (MN/CA IX), CD44v6, Sonic Hedgehog (Shh), Wue-1, plasmacytomas antigen (membrane-bound) IgE, chondroitin sulfate of melanoma proteoglycan (MCSP), CCR8, the precursor of TNF-alpha, STEAP, mesothelin, A33 antigen, stem cell antigen of the prostate (PSCA), Ly-6; desmoglein 4, a neo-epitope of E-Katerina, receptor fetal acetylcholine, CD25, a marker CA19-9 marker CA-125 and receptor P-type Muellerian-inhibiting substance (MIS), sTn (sieciowego antigen, TAG72), FAP (antigen activation of fibroblasts), indocyanine, EGFRvIII, L6, SAS, CD63, TAG72, TF antigen, crustal antigen CD7, CD22, Igα, Igβ, G250, gp100, MT-MMPs, P19 antigen, CO-29 and EphA2.

19. CD3 specific binding construct according to claim 1, in which the specified second binding domain derived from Ig, includes the site of interaction with the antigen having specificity against CCR5.

20. CD3 specific binding construct according to claim 19, in which the CD3 specific binding construct comprises the amino acid is the selected, selected from the group
(a) the amino acid sequence represented by one of the sequences SEQ ID NO.:206, 208, 210, 212, 214 or 216;
(b) amino acid sequence encoded by the nucleic acid sequence represented by one of the sequences SEQ ID NO.:205, 207, 209, 211, 213 or 215; and
(C) the amino acid sequence encoded by a nucleic acid sequence which is degenerate as a result of the genetic code to the nucleotide sequence of (b),
(g) the amino acid sequence encoded by the nucleic acid sequence, hybridities with complementary chain nucleic acid sequence described in (b) under conditions of stringent hybridization and having specificity against CD3 and molecules selected from the group consisting of Arcam, CCR5, CD 19, HER-2, HER-2 neu, HER-3, HER-4. EGFR, PSMA, CEA, MUC-1 (mucin), MUC2, MUC3, MUC4, MUC5AC, MUC5B, MUC7, antigen, Lewis-Y, CD20, CD33, CD30, ganglioside GD3, 9-O-acetyl-GD3, GM2, Globo H, fucosyl GM1, poly SA, GD2, carbonic anhydrase IX (MN/CA IX), CD44v6, Sonic Hedgehog (Shh), Wue-1, plasmacytomas antigen (membrane-bound) IgE, chondroitin sulfate of melanoma proteoglycan (MCSP), CCR8, the precursor of TNF-alpha, STEAP, mesothelin, A33 antigen, stem cell antigen of the prostate (PSCA), Ly-6; desmoglein 4, a neo-epitope of E-Katerina, receptor fetal acetylcholine, CD25, a marker CA19-9, m is rcera CA-125 and receptor type II Muellerian-inhibiting substance (MIS), sTn (sieciowego antigen, TAG72), FAP (antigen activation of fibroblasts), indocyanine, EGFRvIII, L6, SAS, CD63, TAG72, TF antigen, crustal antigen CD7, CD22, Iga, Igβ, G250, gp100, MT-MMPs, P19 antigen, CO-29 and EphA2.

21. CD3 specific binding construct according to claim 1, in which the specified second binding domain derived from Ig, includes the site of interaction with the antigen having specificity against CD 19.

22. CD3 specific binding construct according to item 21, in which the CD3 specific binding construct comprises the amino acid sequence selected from the group
(a) the amino acid sequence represented by one of the sequences SEQ ID NO.:190, 192, 194, 196, 198, 200, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 357, 359,361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 407 or 409;
(b) amino acid sequence encoded by the nucleic acid sequence represented by one of the sequences SEQ ID NO.:189, 191, 193, 195, 197, 199, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394, 396, 398, 400, 402, 404, 406 or 408; and
(C) the amino acid sequence encoded by a nucleic acid sequence which is degenerate as a result of the genetic code to the nucleotide sequence of (b);
(g) amino acid sequences that code is generated by a sequence of nucleic acid, hybridises with complementary chain nucleic acid sequence described in (b), under conditions of stringent hybridization and having specificity against CD3 and molecules selected from the group consisting of Arcam, CCR5, CD 19, HER-2, HER-2 neu, HER-3, HER-4, EGFR, PSMA, CEA, MUC-1 (mucin), MUC2, MUC3, MUC4, MUC5AC, MUC5B, MUC7, antigen, Lewis-Y, CD20, CD33, CD30, ganglioside GD3, 9-O-acetyl-GD3, GM2, Globo H, fucosyl GM1, poly SA, GD2, carbonic anhydrase IX (MN/CA IX), CD44v6, Sonic Hedgehog (Shh), Wue-1, plasmacytomas antigen (membrane-bound) IgE, chondroitin sulfate of melanoma proteoglycan (MCSP), CCR8, the precursor of TNF-alpha, STEAP, mesothelin, A33 antigen, stem cell antigen of the prostate (PSCA), Ly-6; desmoglein 4, a neo-epitope of E-Katerina, receptor fetal acetylcholine, CD25, a marker CA19-9 marker CA-125 and receptor type II Muellerian-inhibiting substance (MIS), sTn (sieciowego antigen, TAG72), FAP (antigen activation of fibroblasts), indocyanine, EGFRvIII, L6, SAS, CD63, TAG72, TF antigen, crustal antigen CD7, CD22, Iga, Igβ, G250, gp100, MT-MMPs, P19 antigen, CO-29 and EphA2.

23. CD3 specific binding construct according to claim 1, in which the specified second binding domain derived from Ig, includes the site of interaction with the antigen having specificity against CD20.

24. CD3 specific binding construct according to item 23, in which the CD3 specific binding construct comprises the amino acid is the selected, selected from the group
(a) the amino acid sequence represented by one of the sequences SEQ ID NO.:218, 220, 222, 224, 226 or 228;
(b) amino acid sequence encoded by the nucleic acid sequence represented by one of the sequences SEQ ID NO.:217, 219, 221, 223, 225 or 227; and
(C) the amino acid sequence encoded by a nucleic acid sequence which is degenerate as a result of the genetic code to the nucleotide sequence of (b).
(g) the amino acid sequence encoded by the nucleic acid sequence, hybridities with complementary chain nucleic acid sequence described in (b), under conditions of stringent hybridization and having specificity against CD3 and molecules selected from the group consisting of Arcam, CCR5, CD 19, HER-2, HER-2 neu, HER-3, HER-4, EGFR, PSMA, CEA, MUC-1 (mucin), MUC2, MUC3, MUC4, MUC5AC, MUC5B, MUC7, antigen, Lewis-Y, CD20, CD33, CD30, ganglioside GD3, 9-O-acetyl-GD3, GM2, Globo H, fucosyl GM1, poly SA, GD2, carbonic anhydrase IX (MN/CA IX), CD44v6, Sonic Hedgehog (Shh), Wue-1, plasmacytomas antigen (membrane-bound) IgE, chondroitin sulfate of melanoma proteoglycan (MCSP), CCR8, the precursor of TNF-alpha, STEAP, mesothelin, A33 antigen, stem cell antigen of the prostate (PSCA), Ly-6; desmoglein 4, a neo-epitope of E-Katerina, receptor fetal acetylcholine, CD25, a marker CA19-9, is arcara CA-125 and receptor type II Muellerian-inhibiting substance (MIS), sTn (sieciowego antigen, TAG72), FAP (antigen activation of fibroblasts), indocyanine, EGFRvIII, L6, SAS, CD63, TAG72, TF antigen, crustal antigen CD7, CD22, Igα, Igβ, G250, gp100, MT-MMPs, P19 antigen, CO-29 and EphA2.

25. Nucleic acid encoding a CD3 specific binding construct according to one of claims 1 to 24.

26. The vector for the expression of CD3-specific connecting structure according to any one of claims 1 to 24, comprising a nucleic acid according A.25.

27. Vector on p which further includes nucleic acid, which is a regulatory nucleic acid is functionally linked to a specific nucleic acid according A.25.

28. Vector on p, in which the vector is expressing vector.

29. Eukaryotic a host cell transformed or transfectional vector according to one of PP-28.

30. A method of obtaining a CD3-specific connecting structure according to one of claims 1 to 24; this method includes culturing the host under clause 29 under conditions that allow expression of the polypeptide structure and the allocation of the formed polypeptide constructs of culture.

31. Composition for treatment or prevention of proliferative diseases, comprising an effective amount of CD3-specific connecting structure according to one of claims 1 to 24, and, optionally, the protein link is, able to provide an activating signal for immune effector cells.

32. Composition for preventing, treating or alleviating proliferative diseases, neoplastic diseases, inflammatory diseases, immunological disorders, infectious disease, viral disease, allergic reactions, parasitic reactions, graft versus host" or "reaction of the host against the graft, including an effective amount of CD3-specific linking structure obtained by the method according to item 30, and, optionally, a protein compound capable of activating signal for immune effector cells.

33. Composition for preventing, treating or alleviating proliferative diseases, neoplastic diseases, inflammatory diseases, immunological disorders, infectious disease, viral disease, allergic reactions, parasitic reactions, graft versus host" or "reaction of the host against the graft, including an effective amount of CD3-specific binding design obtained using nucleic acid A.25, and, optionally, a protein compound capable of activating signal for immune effector cells.

34. Composition for prevention, treatment is or alleviating proliferative diseases, neoplastic diseases, inflammatory diseases, immunological disorders, infectious disease, viral disease, allergic reactions, parasitic reactions, graft versus host" or "reaction of the host against the graft, including an effective amount of CD3-specific binding design obtained using the vector according to one of p-28 and, optionally, a protein compound capable of activating signal for immune effector cells.

35. Composition for preventing, treating or alleviating proliferative diseases, neoplastic diseases, inflammatory diseases, immunological disorders, infectious disease, viral disease, allergic reactions, parasitic reactions, graft versus host" or "reaction of the host against the graft, including an effective amount of CD3-specific linking structure obtained by the owner on clause 29 and, optionally, a protein compound capable of activating signal for immune effector cells.

36. The composition according to p, which is a pharmaceutical composition, optionally optionally comprising suitable formulations of carrier, stabilizers and/or excipients, or diagnostic component is izia, optional optional including means and methods of detection.

37. The composition according to p, which is a pharmaceutical composition, optionally optionally comprising suitable formulations of carrier, stabilizers and/or excipients, or diagnostic composition, optionally optionally including means and methods of detection.

38. The composition according to p, which is a pharmaceutical composition, optionally optionally comprising suitable formulations of carrier, stabilizers and/or excipients, or diagnostic composition, optionally optionally including means and methods of detection.

39. The composition according to clause 34, which is a pharmaceutical composition, optionally optionally comprising suitable formulations of carrier, stabilizers and/or excipients, or diagnostic composition, optionally optionally including means and methods of detection.

40. The composition according to p, which is a pharmaceutical composition, optionally optionally comprising suitable formulations of carrier, stabilizers and/or excipients, or diagnostic composition, optionally optionally including means and methods of detection.

41. CD3-specific connecting structure according to one of claims 1 to 24 for warnings, cured the I or alleviating proliferative diseases, neoplastic diseases, inflammatory diseases, immunological disorders, infectious disease, viral disease, allergic reactions, parasitic reactions, graft versus host" or "reaction of the host against the graft".

42. CD3-specific linking structure obtained by the method according to item 30, for preventing, treating or alleviating proliferative diseases, neoplastic diseases, inflammatory diseases, immunological disorders, infectious disease, viral disease, allergic reactions, parasitic reactions, graft versus host" or "reaction of the host against the graft".

43. CD3 specific binding construct, obtained using nucleic acid A.25, for preventing or alleviating proliferative diseases, neoplastic diseases, inflammatory diseases, immunological disorders, infectious disease, viral disease, allergic reactions, parasitic reactions, graft versus host" or "reaction of the host against the graft".

44. CD3 specific binding construct, obtained using the vector according to one of p-28 to prevent, treat or alleviate a proliferative disease, a tumorous the th disease, inflammatory diseases, immunological disorders, autoimmune disease, infectious disease, viral disease, allergic reactions, parasitic reactions, graft versus host" or "reaction of the host against the graft".

45. CD3-specific linking structure obtained by the owner on clause 29, to prevent, treat or alleviate a proliferative disease, a tumorous disease, an inflammatory disease, an immunological disorder, an autoimmune disease, infectious disease, viral disease, allergic reactions, parasitic reactions, graft versus host" or "reaction of the host against the graft".

46. A method of preventing, treating or alleviating proliferative disease, a tumorous disease, an inflammatory disease, an immunological disorder, an autoimmune disease, infectious disease, viral disease, allergic reactions, parasitic reactions, graft versus host" or "reaction of the host against the graft, including the introduction of CD3-specific connecting structure according to claims 1-24 to a subject in need of such prevention, treatment or relief.

47. Method of prevention, treatment or obleceni the proliferative disease, neoplastic diseases, inflammatory diseases, immunological disorders, autoimmune disease, infectious disease, viral disease, allergic reactions, parasitic reactions, graft versus host" or "reaction of the host against the graft, including the introduction of CD3-specific linking structure obtained by the method according to item 30, to a subject in need of such prevention, treatment or relief.

48. A method of preventing, treating or alleviating proliferative disease, a tumorous disease, an inflammatory disease, an immunological disorder, an autoimmune disease, infectious disease, viral disease, allergic reactions, parasitic reactions, graft versus host" or "reaction of the host against the graft, including the introduction of CD3-specific binding design obtained using nucleic acid A.25, to a subject in need of such prevention, treatment or relief.

49. A method of preventing, treating or alleviating proliferative disease, a tumorous disease, an inflammatory disease, an immunological disorder, an autoimmune disease, infectious disease, viral disease, allergic reactions, parasite is ical reactions, graft versus host" or "reaction of the host against the graft, including the introduction of CD3-specific binding design obtained using the vector according to one of PP-28, to a subject in need of such prevention, treatment or relief.

50. A method of preventing, treating or alleviating proliferative disease, a tumorous disease, an inflammatory disease, an immunological disorder, an autoimmune disease, infectious disease, viral disease, allergic reactions, parasitic reactions, graft versus host" or "reaction of the host against the graft, including the introduction of CD3-specific linking structure obtained by the owner under clause 29 to a subject in need of such prevention, treatment or relief.

51. The method according to item 46, in which the specified subject is people.

52. The method according to p in which the specified subject is people.

53. The method according to p in which the specified subject is people.

54. The method according to § 49, in which the specified subject is people.

55. The method according to item 50, in which the specified subject is people.

56. The method according to item 46, further including the introduction of protein compounds capable of activating signal for immune effectivities.

57. The method according to p, optionally including the introduction of protein compounds capable of activating signal for immune effector cells.

58. The method according to p, optionally including the introduction of protein compounds capable of activating signal for immune effector cells.

59. The method according to § 49, further including the introduction of protein compounds capable of activating signal for immune effector cells.

60. The method according to item 50, further including the introduction of protein compounds capable of activating signal for immune effector cells.



 

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7 cl, 12 dwg, 1 tbl, 3 ex

FIELD: chemistry.

SUBSTANCE: proposed is a recombinant single-strand trispecific antibody for treating tumours which express CEA. The said antibody consists of a series of three antibody fragments: anti-CEA-scFv, anti-CD3-scFv and VH CD28-antibody, linked by two intermediate linkers (intermediate linker Fc and intermediate linker HSA). If necessary, a c-myc-mark or (His)6-mark can be added at the C-end. Described is DNA, which codes the antibody, expression vector based on it and E.coli cell, containing the vector.

EFFECT: use of the invention is more beneficial in clinical use compared to bispecific antibodies and known trispecific antibodies, makes easier clearing and expression of an antibody, which can further be used in treating CEA-mediated tumours.

10 cl, 21 dwg, 11 ex

FIELD: biotechnology.

SUBSTANCE: present invention relates to biotechnology and immunology. Proposed here is a polynucleotide, encoding a cyclic single-stranded tri-specific antibody. The antibody is directed against human ovarian carcinoma in vitro, has mass of approximately 84 kD and consists of three components: an antibody against human ovarian carcinoma cells, anti-CD3 antibody and anti-CD28 antibody, which are joined together by peptide interlinks such that, they form a cyclic antibody. Invented is an expression vector, containing a coding polynucleotide and versions of E.coli host cell based on the polynucleotide and expression vector.

EFFECT: use of the invention provides for a stable antibody molecule, optimum for activation of T-cells, which can be used in curing human ovarian carcinoma.

8 cl, 12 dwg

FIELD: medicine.

SUBSTANCE: versions of the bond intended for linkage with the external domain B (ED-B) of a fibronectin are offered. The bond includes an antigen-binding fragment of one-chained antibody L19 and a cysteinum-containing linker for hanging of a radioactive label. Versions of a pharmaceutical composition for diagnostics and treatment of angiogenic diseases on the basis of the specified bond are opened. Application of bond for linkage with radioactive bond is described. The method of reception of bond in eucariotic cells is opened, including in Pichia pastoris and a set for reception is radioactive labelled agents on the basis of bond.

EFFECT: high-avid bond accumulation in solid tumours.

23 cl, 4 dwg, 5 tbl, 15 ex

FIELD: chemistry, biochemistry.

SUBSTANCE: current invention relates to the field of biotechnology and immunology. Proposed is an antibody, specific to the human ED-B. Antibody specified is a molecule in the form of either dimerizated mini-immunoglobulin or IgG1, whose variable region comes from the antibody L19. In case the mini-immunoglobulin variable region L19 is merged with εS2-CH4, then as in the case IgG1, the variable region L19 is merged with the constant domain of IgG1. Conjugates of antibodies with radioisotopes have been discovered. Described is the coding nucleic acid, carrying its host cell, capable of producing antibodies, and method of obtaining antibodies from cells. Discovered is a method of determining the degree of bonding of antibodies, also compositions based on antibodies. Described is the use of antibodies for preparing medicine for treating either damage related to angiogenesis, or for treating tumours. Utilisation of the invention provides antibodies, which possess high accumulating capacity to tumours, improved capability to bonding with radioactive labels and unexpectedly retains immunoreactivity in the plasma, in comparison to scFv L19. Antibody specified can be used in diagnostics and treatment of tumours.

EFFECT: obtaining antibodies which can be used in diagnostics and treatment of tumours.

22 cl, 13 dwg, 8 tbl

FIELD: medicine, microbiology.

SUBSTANCE: invention concerns biotechnology. It is described bispecific antibody which binds also the factor of blood coagulation IX or the activated factor of blood coagulation IX, and the factor of blood coagulation X, and functionally replaces the factor of blood coagulation VIII or the activated factor of blood coagulation VIII which strengthens enzymatic reaction. The pharmaceutical composition containing the described antibody is revealed. The present invention can be used as an alternative agent for functional replacement of cofactor which strengthens enzymatic reaction.

EFFECT: creation of bispecific antibody which can replace functional proteins, strengthens enzymatic reaction.

14 cl, 18 dwg, 37 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology and immunology. Claimed is therapeutically active fused protein with reduced immunogenicity. Protein consists of two proteins derived from human proteins connected through the fusion region. Connective region, which covers or surrounds fusion region within the limits from 1 to 25 amino acid residues, contains modification, which removes T-cell epitope, in norm absent in humans. Claimed is application of fused protein for obtaining pharmaceutical composition for tumour treatment. Claimed is nucleic acid coding fused protein. Method of reduction of fused protein immunogenicity by introduction of substitutes of corresponding amino acids is described. Application of the invention allows reducing ability of connective epitope of therapeutically active fused protein to bind with molecules of the main complex of hystocompatibility (MHC) of class II, which finally reduces interaction of epitope with receptors of T-cells and can find application in medicine for prevention of immunological disorders arising with introduction of therapeutically active protein non-modified in connective region.

EFFECT: reduction of interaction of epitope with receptors of T-cells, which can find application in medicine for prevention of immunological disorders arising with introduction of therapeutically active protein non-modified in connective region.

23 cl, 12 ex

FIELD: bioengineering.

SUBSTANCE: versions of the molecule binding CD45RO and CD45RB, and the anti-CD45RO and anti-CD45RB antibody are invented. In one of versions, the said molecule contains at least one antigen-binding site and includes the subsequently located hypervariable sites CDR1, CDR2 and CDR3. The molecule represents the humanised or monoclonal antibody. CDR1 has the amino acid sequence NYIIH, CDR2 has the amino acid sequence YFNPYNHGTKYNEKFKG and CDR3 has the amino acid sequence SGPYAWFDT. The molecule can additionally contain the subsequently located hypervariable sites CDR1', CDR2' and CDR3'. CDR1' has the amino acid sequence RASQNIGTSIQ, CDR2' has the amino acid sequence SSSESIS and CDR3' has the amino acid sequence QQSNTWPFT. In another version, the molecule contains both heavy and light chains where the amino acid sequences contain the corresponding CDR. The versions of the corresponding coding polynucleotide are disclosed; expression vector and based on it expression system. The host cell is disclosed basing on the expression system. The application of the molecule in treatment of autoimmune diseases, graft rejection, psoriasis, intestine inflammatory disease and allergy is described. The pharmaceutical composition for the said application is disclosed.

EFFECT: enables immunosuppressant induction; inhibiting T-cell response and primary lymphocyte response in mixed lymphocyte culture (MLC); prolongs survival period in mice with severe combined immunodeficiency SCID.

20 cl, 5 dwg, 2 tbl, 8 ex

FIELD: biotechnology, immunology.

SUBSTANCE: disclosed are variants of chimerical anti-IL-6 antibodies based on mice CLB-8 antibody. Each antibody contains constant region from one or more human antibodies. Described are variants of nuclear acids encoding anti-IL-6 antibody, vectors and host cells. Developed is method for production of anti-IL-6 antibody by using nuclear acid or vector. Described are variants of composition for application in method for modulation of malignant disease or immune disorder mediated with IL-6. Developed is method for treatment or modulation of malignant disease or immune disorder mediated with IL-6.

EFFECT: variant of chimerical anti-IL-6 antibody with high affinity of mice anti-IL-6 antibody and reduced immonogenicity.

26 cl, 16 dwg, 1 tbl, 8 ex

FIELD: chemistry; biochemistry.

SUBSTANCE: invention relates to biotechnology and is a tumour necrosis factor receptor 1 antagonist which is a TNFR1 domain antibody (dAb). The invention also pertains to a TNFR1 domain antibody (dAb) monomer which binds the tumour necrosis factor receptor 1, as well as to a ligand which contains such a monomer. The invention also relates to a method of inhibiting signal transmission through the TNFR1 receptor using said antagonists. The invention also discloses nucleic acid which codes said antagonist, a vector containing said nucleic acid, a host cell containing said vector, a pharmaceutical composition containing said antagonist or said dAb monomer.

EFFECT: invention can be effective in treating inflammatory diseases (for example chronic inflammatory diseases).

57 cl, 99 dwg, 14 tbl, 20 ex

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