Anti-cd40 antibody mutants

FIELD: pharmacology.

SUBSTANCE: present invention refers to immunology and biotechnology. There are antibody-antagonist to CD40 with their variable areas derived from an antibody produced of hybridoma 4D11 (FERM BP-7758). The constant areas of antibodies are derived from human IgG4 with mutations S228P and L235E. There are described related coding polynucleotides and the based expression vector. There is disclosed host-cell containing said vector. There is described method for preparing monoclonal antibody and application thereof in the pharmaceutical composition.

EFFECT: application of the invention provides reduced ADCC and CDC activity that can find application in therapy of autoimmune diseases and graft rejection.

10 cl, 26 dwg, 2 tbl, 22 ex

 

The text descriptions are given in facsimile form.

1. Monoclonal antibody binding to human CD40, comprising (i) two heavy chains, each of which contains a constant region derived from human IgG4, with the substitution of serine with Proline at position 228 and the substitution of LEU is in glutamic acid at position 235, and the variable region of the heavy chain of the monoclonal antibody produced by hybridoma 4D11 (no access FERM BP-7758), and (ii) two light chains, each of which contains the variable region of the light chain of the monoclonal antibodies produced by hybridoma 4D11 (no access FERM BP-7758).

2. Monoclonal antibody binding to human CD40, comprising (i) two heavy chains, each of which contains a constant region derived from human IgG4, with the substitution of serine with Proline at position 228 and the substitution of leucine glutamic acid at position 235, and variable region represented by amino acid sequence in the range from Q at position 27 to S at position 147 in SEQ ID NO:46, and (ii) two light chains, each of which contains a variable region represented by amino acid sequence in the range from position 23 To position 128 in SEQ ID NO:48.

3. Monoclonal antibody binding to human CD40, comprising two heavy chains, each of which presents the amino acid sequence in the range from Q at position 27 To position 474 in SEQ ID NO:140, and two light chains, each of which presents the amino acid sequence in the range from position 23 to position 235 in SEQ ID NO:142.

4. Polynucleotide represented by SEQ ID NO:139, to obtain antibodies in which the yubom one of claims 1 to 3.

5. Polynucleotide represented by a sequence in the range from position 79 to position 1425 in SEQ ID NO:139, to obtain the antibody according to any one of claims 1 to 3.

6. The expression vector containing polynucleotide represented by SEQ ID NO:139, and polynucleotide represented by SEQ ID NO:141, to obtain the antibody according to any one of claims 1 to 3.

7. A host cell containing the expression vector of claim 6.

8. The method of obtaining monoclonal antibodies, providing the stage of culturing the host cell according to claim 7 in culture medium and obtaining the monoclonal antibody from the culture and/or host.

9. Pharmaceutical composition containing an effective dose of a monoclonal antibody according to any one of claims 1 to 3 as an active ingredient for prevention or treatment of transplant rejection, autoimmune diseases, allergies or inhibition of factor VIII clotting.

10. The use of monoclonal antibodies according to any one of claims 1 to 3 to obtain a pharmaceutical composition for the prevention or treatment of transplant rejection, autoimmune diseases, allergies or inhibition of factor VIII clotting.



 

Same patents:

FIELD: medicine.

SUBSTANCE: method is suggested for production of antibody for binding to NK-cells, which crossly interacts with products of gene KIR2DL1 and KIR2DL2/3 and neutralises inhibitor activity of such KIR. Mentioned method includes selection of such antibodies that crossly interact at least with products of gene KIR2DL1 and KIR2DL2/3, are able to restore lysis with NK cells Cw3+ or Cw4+ target cells and are bound with NK cells or polypeptide of KIR primate. Antibodies produced by this method are described, as well as their derivatives, where antibody is linked with toxin, radionuclide, recognisable aggregation, solid carrier or polyethylene glycol.

EFFECT: invention provides for preparation of single type of antibodies, which controls activity of NK cells of various type, provides for amplification of their cytotoxicity, which may find application in therapy, for increase of activity or cytotoxicity of NK cells in individuals without preliminary detection of HLA type in individual.

7 cl, 13 dwg, 4 tbl, 7 ex

FIELD: medicine.

SUBSTANCE: invention refers to antibody specifically getting bound with PRO87299 version. In addition, the antibody according to the invention has ability to block interaction HVEM and PRO87299 and to function as PRO87299 agonist. The antibody of agonist nature is produced by hybridoma Btig5F5.1 or Btig3B1.9. For the antibody, there is established amino acid sequence given in the description. The invention discloses the methods of using the antibodies to stimulate or reduction of immune response in immune-associated diseases connected, to relieve lymphoma, and inflammatory disease in requiring mammal, to detect polypeptide PRO87299 in a sample and to manage rejection of grafted cells.

EFFECT: antibody is an immunomodulator that allows applying therapeutically identical medicinal agents both to intensify and reduce immune response.

16 cl, 34 dwg, 7 tbl, 20 ex

FIELD: medicine.

SUBSTANCE: strain A-4A7 is prepared by fusion of mouse myeloma cells of line SP2/0.Agl4 with mouse lymphocytes of line Balb/c immunised by introduction in pads of a purified preparation AMGF (alpha2-microglobulin of fertility) separated from amniotic fluid, and deposited in the transplantable mammal cell culture collection of the Research Institute of Human morphology of the Russian Academy of Medical Science numbered 131/2002. Strain A-4A7 synthesises monoclonal antibodies (MCA) of IgGI class specifically reacting in solid-phase immune-enzyme analysis (IEA) with AMGF isoforms of endometrial, follicular and sperm nature. Activity of the strain: cultural supernatant contains MCA 3-5 mkg/ml, while ascetic fluid contains MCA 2-5 mg/ml. The antibody titre in cultural fluid is 1:500-1:1000, in ascetic fluid up to 1:1×107. A-4A7 bonds various AMGF protein glycoforms produced in male and female reproductive organs. Application of MCA A-4A7 as an immunodiagnosis test systems allows for high-specific and high-sensitive (1 ng/ml) quantitative analysis of various AMGF/glykodeline isoforms in biological liquids.

EFFECT: new compounds are characterised with valuable biological properties.

2 ex

FIELD: medicine.

SUBSTANCE: according to the present invention, there are disclosed disease diagnostic techniques and sets characterised by nonphysiological content of protein Hepsidin in mammals. Substance of the invention lies in binding antibody and polypeptide, where antibody or its fragment is specifically bound with one or more average epitopes or epitopes of carboxyl termination of sequence SEQ ID NO: 2. Additionally, there are disclosed relevant antibodies.

EFFECT: disease screening characterised by nonphysiological content of protein Hepsidin in organism.

19 cl, 2 ex, 16 dwg, 2 tbl

FIELD: medicine.

SUBSTANCE: method for making polyclonal Nogo protein antibodies by protein immunisation of an animal, with at least 6 amino-acid residues of active area Nogo A, free from any other myelin material of central nervous system whereto bound in natural conditions. Said active area covers position 1-171, 260-974, 1163-1178 of the corresponding amino acid sequence of protein Nogo A. There is disclosed separated antiserum based on polyclonal antibodies made under said method. There are disclosed methods for immunisation of a non-human animal to make polyclonal antibodies, as well as method for making monoclonal antibody, and the specified monoclonal antibody.

EFFECT: making active protein Nogo A antibodies to be applied in medicine for neurite growth activation.

26 cl, 18 dwg, 2 tbl, 8 ex

FIELD: biology.

SUBSTANCE: present invention relates to biotechnology. An A-5D1 strain is obtained from fusing mouse myeloma cell line SP2/0.Ag14 with mouse lymphocyte line Balb/c, immunised by introduction into pouches of a purified preparation of alpha 2-fertility microglobulin, extracted from amniotic fluid, and deposited in the collection of passaged culture of mammal cells of the state research institute of human morphology (Russian Academy of Medical Sciences) under the number 144/2002. The strain of hybredome A-5D1 synthesises the IgG1 class monoclonal antibody, which specifically interacts in enzyme-linked immunosorbent assay and immunohistochemical assay with isoforms of alpha 2-fertility microglobulin of endometrial or special origin. Monoclonal antibodies do not have cross reaction with placental α 1-microglobulin, trophoblastic β1-globulin, human chorionic gonadotropin, α-fetoprotein, bovine serum albumin, and C-reactive protein. Monoclonal antibodies detect alpha 2-fertility microglobulin/glycodelin in cells and tissue of female and male reproductive organs.

EFFECT: use of monoclonal antibody A-5D1 as an immunodiagnostic system allows for quantitative analysis of isoforms of alpha 2-fertility microglobulin/glycodelin in body fluids with high sensitivity (1 ng/ml) and specificity.

3 ex

FIELD: medicine; pharmacology.

SUBSTANCE: allocated human monoclonal antibodies which specifically bind a receptor of the epidermal growth factor (EGFR), and also corresponding compositions on the basis of antibodies and a biospecific molecule are described. Human antibodies can be received with use of the transgenic mouse capable to formation of set of isotypes of human monoclonal antibodies by recombination V-D-J and switching of isotypes. The pharmaceutical compositions containing human antibodies for treatment or prevention of diseases, mediated by expression EGFR, the transgenic animals distinct from a human, the specified expressing antibodies, hybridomes and transfectomes which produce human antibodies are also presented. Ways of therapy and diagnostics of the diseases mediated by expression EGFR, with use of human antibodies or their antigen-binding of fragments, and also methods of growth suppression of the cells expressing EGFR, and an induction of cytolysis of the specified cells are described.

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53 cl, 22 dwg, 4 tbl, 11 ex

FIELD: biotechnologies.

SUBSTANCE: obtaining monoclonal antibodies immunoreactive with the nucleocapsid protein of the hepatitis virus C-4G5, and the development of a method of diagnostics based on the double layer version of immunoenzymometric assay for the detection of the nucleocapsid (core protein) of the hepatitis C virus using a combination of linear and conformational core protein epitope monoclonal antibodies 27, 1F9, and 4G5: MCA 1F9: linear epitope with amino-acid residues (AAR) 81-85, MCA 27, and 4G5: conformational epitopes at site 1-150 of the core protein AAR. The combination of monoclonal antibodies 27, 1F9, and 4G5, immunoreactive with the hepatitis C virus nucleocapsid protein may be used both for the capture (capture antibodies) and for the detection (detection antibodies) of the core protein. The double layer variant of the immunoenzymometric assay developed for the detection of the nucleocapsid of the hepatitis C virus using a combination of monoclonal antibodies including MCA 4G5 has high sensitivity and allows to detect core proteins in blood of both asymptomatic HCV-infected donors and patients suffering from acute and chronic hepatitis C. The advantages of the immunoenzymometric assay of the core protein may be useful for determining the viral load during IF therapy.

EFFECT: allows to diagnose hepatitis C with high probability at early stages of disease and is efficient for detection of HCV of at least three genotypes most actual for Russia.

3 cl, 3 tbl, 4 ex

FIELD: medicine, peptides, biochemistry, pharmacy.

SUBSTANCE: invention relates to modification of glycosylation of proteins for preparing polypeptides with improved therapeutic indices including antibodies with enhanced antibody-dependent cellular cytotoxicity. For preparing indicated polypeptides cell-host is used that is modified with nucleic acid encoding enzyme β-1,4-N-acetylglucosaminyltransferase III (GnTIII). Prepared polypeptide represents, in particular, IgG or its fragment. Invention discloses a method for preparing polypeptide and antibodies or its fragment and a fusion protein prepared by indicated method. Invention describes a pharmaceutical composition used for increasing Fc-mediated cellular cytotoxicity and comprising antibody and carrier, and its using in cancer treatment, and a method for treatment of disease associated with elevated amount or production of B cells using indicated antibody, in particular, against CD20, and representing antibody IDEC-C2B8 in the preferable variant. Invention provides preparing polypeptide and antibody possessing the enhanced Fc-mediated cellular cytotoxicity that decrease the content of B cells in a patient body.

EFFECT: improved preparing method, valuable medicinal properties of polypeptide and antibodies.

38 cl, 21 dwg, 4 ex

FIELD: biotechnology, immunology.

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EFFECT: valuable biological and medicinal properties of antibodies.

53 cl, 4 tbl, 10 dwg, 2 ex

FIELD: pharmacology.

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35 cl, 13 dwg, 36 tbl, 14 ex

FIELD: medicine.

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16 cl, 3 tbl, 7 ex

FIELD: food industry.

SUBSTANCE: strain Streptococcus thermophilus which produces lactic acid is described. Sequence of nucleic acids made of the strain producing polysaccharides are also described as well as food or pharmaceutical composition and milk product containing such strain.

EFFECT: strain has strong structural properties.

16 cl, 4 dwg, 6 tbl, 5 ex

FIELD: medicine.

SUBSTANCE: invention relates to field of genetic engineering and medicine. Described is animal, non-human, having sequence of nucleic acid encoding presenilin 1, carrying mutations, corresponding to M233T and L235P mutations in PS1 protein of mouse. Animal also contains sequence of nucleic acid, encoding whole gene or part of gene, encoding APP. APP protein represents APP751, originates from human and carries mutations Swedish and London. Animal is intended for application in fight against Alzheimer's disease. Also described are PS1 protein and encoding it nucleic acid.

EFFECT: invention can be used in medicine for discovering compounds intended for Alzheimer's disease treatment.

20 cl, 50 dwg, 1 tbl, 8 ex

FIELD: chemistry; biochemistry.

SUBSTANCE: invention relates to biotechnology, specifically to production of versions of the Gla domain of human factor VII or human factor VIIa, and can be used in medicine. Amino acid sequence of the FVII or FVIIa version is obtained, which differs on 1 to 15 amino acid residues with amino acid sequence of the human factor VII (hFVII) or human factor VIIa (hFVIIa), in which a negatively charged amino acid residue is introduced by substitution in position 36. Obtained variants of FVII or FVIIa are used in a composition for treating mammals with diseases or disorders, where blood clotting is desirable.

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42 cl, 3 dwg, 5 tbl, 11 ex

FIELD: medicine.

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EFFECT: invention allows for making gamma-carboxylated vitamin K dependent protein in production quantities.

29 cl, 5 dwg, 6 tbl, 7 ex

FIELD: medicine.

SUBSTANCE: invention claims synthetic DNS molecule encoding L1 HPV58 protein, where codons are optimised for high expression level in yeast cell. Also invention claims expression vector, yeast host cell, HPV58 virus-like particle and method of its obtainment, and pharmaceutical composition including such VLP.

EFFECT: invention can be applied in medicine for efficient immune prevention of papillomavirus infection by neutralising antibodies and cell-mediated immunity, and for treatment of developed HPV infections.

10 cl, 10 dwg, 8 ex

FIELD: medicine.

SUBSTANCE: invention includes obtaining and applying the versions of allergens of 5 Pooideae group, which are characterised by decreased IgE reactivity in comparison to known wild-type allergens and at the same time by essentially retained reactivity in comparison to T-lymphocytes. Versions of allergens, as per the invention, do not have at least one section or a combination of sections corresponding to amino-acid sequence of sections Phl p5a 94-113 or 175-198, which is given in the description. Versions of allergens have been obtained by gene-engineering methods. In the invention is opened DNA molecule encoding the version of allergen, recombinant expression vector, host organism, expression version of allergen and method of obtaining the version of allergen by using the above described means. Such hypoallergic versions of allergens can be used as a remedy against allergies determined by allergens of 5 Pooideae group for specific immunotherapy (hyposensitisation) of the patients having grass pollen allergy or for preventive immunotherapy of grass pollen allergies by using pharmaceutical composition.

EFFECT: preparations based on versions of allergens, as per the invention, have decreased IgE reactivity and the retained reactivity in relation to T-lymphocytes.

12 cl, 17 dwg, 3 tbl

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: chemistry; medicine.

SUBSTANCE: claimed are polypeptide and respective polynucleotide zcytor17lig and molecules of antibody against human zcytor17. Human zcytor17lig is novel cytokine. Claimed invention also relates to methods of protein obtaining, its application for stimulation of immune reaction in mammal. Described is method of obtaining antibodies to said protein and respective antibodies.

EFFECT: polypeptides can be used in realisation of methods stimulation of immune system, proliferation and development of hemopoietic cells in vitro and in vivo.

17 cl, 3 dwg, 21 tbl, 47 ex

FIELD: pharmacology.

SUBSTANCE: invention concerns immunology and biotechnology. There is offered human monoclonal antibody specific to TNF-alpha containing light and heavy chain with appropriate CDR3 sites. There are described versions thereof including those based on heavy and light chains and coded by human genes VH3-33 and A30VK1 or VH3-53 and L2VK3 respectively. There are disclosed: the method for estimating the TNF-alpha content in the patient's sample with using specified antibodies, and application of antibodies for preparing a medical product. There are described: compositions for diagnostics and treatment of the conditions associated with TNF-alpha activity on the basis of antibodies. There is disclosed coding nucleic acid, a cell for making said antibodies and the method for making said antibodies.

EFFECT: application of the invention ensured high-affinity neutralizing monoclonal antibodies with improved Kd and IC50 in comparison with Infliximab, Adalimumab or Etanercept that can find application in medicine for treatment and diagnostics of the diseases associated with TNF-alpha hyperactivity.

35 cl, 13 dwg, 36 tbl, 14 ex

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