Method for high-efficiency collection of functional cells in vivo
SUBSTANCE: present group of inventions relates to biotechnology and methods of collecting functional cells (versions). The described solutions comprise implantation of an implantable medical vessel under the skin for not more than two weeks, where a cell population is immobilised in the vessel using any of the proteins HMGB1, HMGB2, HMGB3, S100A8, S100A9 or hyaluronic acid or mixtures of any two or more thereof. Said factors have an activity of attracting specific functional cells into the body.
EFFECT: present inventions provide efficient and safe collection of biologically functional cells, particularly stem cells.
7 cl, 37 dwg, 1 tbl, 12 ex
SUBSTANCE: nutrient medium comprises sodium chloride, potassium chloride, magnesium sulphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, calcium chloride, sodium bicarbonate, L-arginine, L-glutamine, L-tyrosine, L-tryptophan, calcium pantothenate, pyridoxal HCl, thiamine, meso-inositol, nicotinamide, riboflavin, folic acid, glucose, enzymatic hydrolyzate of muscle proteins, lactalbumin enzymatic hydrolyzate, sodium salt of benzylpenicillin with activity of 0.09×105-1.1×105, kanamycin sulphate with activity of 0.09×105-1.1×105, nystatin with activity of 6×104 to 6.5×104 IU/l, choline chloride, succinic acid and distilled water in a predetermined ratio of components.
EFFECT: invention enables to improve the quality of the product obtained.
1 tbl, 1 ex
SUBSTANCE: invention refers to biotechnology. What is described is using specific conjugate molecules, which are cargo molecule transporters for transporting a substance of interest into leukocytes. There are described a method for producing the above conjugate molecules, which are the cargo molecule transporters, a method for transporting the substance of interest into leukocytes, and the leukocytes containing the above conjugate molecules, which are the cargo molecule transporters. The invention can be used in medicine.
EFFECT: above conjugate molecules, which are the cargo molecule transporters, can be used for treating, preventing, relieving and/or reducing the intensity of a disease and/or a disorder involving leukocytes.
26 cl, 34 dwg, 5 tbl, 29 ex
SUBSTANCE: blood is separated into plasma and cellular fraction. Then the blood cellular fraction is subjected to successive two-phase treatment first by the buffer PBS solution containing 5 mM of EDTA with subsequent centrifugation and collection of supernatant. Cells are treated by equal volume of 0.15-0.35% trypsin solution in PBS with subsequent centrifugation and collection of supernatant. Plasma and extracted supernatants from cellular fraction are merged, cellular debris is removed by centrifugation at 15000-17000 g within 10-20 minutes. Foreign particles of non-exosome origin by filtration through filters with the diameter pores 0.1 mcm, and the total pool of exosomes is settled by ultracentrifugation at 100000-160000 g during 60-120 minutes.
EFFECT: invention allows to increase yield and purity of target product, reduce duration and labour input of the method.
3 cl, 2 dwg, 1 tbl, 3 ex
SUBSTANCE: strain of cells of the Chinese hamster ovaries CHO-Inflix 20/5 is produced, transfected by vectors, expressing light and heavy chains of chimeric antibody against tumour necrosis factor alpha (TNF-α) of human being is produced. The strain is deposited into the Russian collection of cellular cultures under No. RKKK(P)760D.
EFFECT: invention allows to produce chimeric antibody with the specific efficiency no less than 33,5 picograms per cell a day.
4 dwg, 2 tbl, 4 ex
SUBSTANCE: invention relates to field of biotechnology. Claimed is method of cultivating epithelial stem cells or separated tissue fragments, containing said epithelial stem cells, including provision of extracellular matrix, incubation of epithelial stem cell or separated tissue fragment, containing said epithelial stem cells, with extracellular matrix, cultivation of epithelial stem cell or separated tissue fragment in presence of cell culture medium, which contains basal medium for animal or human cells, into which inhibitor of bone morphogenetic protein (BMP) and 5-500 ng/ml of mitogenic growth factor are added, as well as cell culture medium, its application, method of three-dimensional organoid cultivation, method of its obtaining, three-dimensional organoid and its application.
EFFECT: method of cultivating epithelial stem cells is claimed.
32 cl, 10 ex, 34 dwg
SUBSTANCE: invention relates to biotechnology. Method includes adhesion of cell population to microcarrier in medium, containing Pho-kinase inhibitor. After that, cultivation of cells, separation of cells from microcarrier and adhesion of obtained cells to second microcarrier in medium containing inhibitor of Rho-kinase. Invention can be used in medicine.
EFFECT: method of growing human embryonic stem cells is disclosed.
7 cl, 48 dwg, 3 tbl, 11 ex
SUBSTANCE: invention refers to biotechnology and medicine. What is presented is a method for differentiation of mesenchymal stem cells of fibrotic lungs into fibroblastic cells, characterised by recovering the cells self-maintaining in vitro for 60 days and having mesenchymal stem cell phenotype (CD44+, CD73+, CD90+, CD106+, CD31-, CD34-, CD45-) from the right lobe of fibrotic lungs and producing the fibroblastic cells when adding fibroblast growth factor 2 mcg/ml to the mesenchymal stem cell culture.
EFFECT: method enables producing the donor cells from the patients suffering idiopathic fibrosis from fibrotic pulmonary tissues.
3 dwg, 1 ex
SUBSTANCE: invention relates to the field of biotechnology. ENA-21/13-13-finite monolayer-suspension cell subline of kidney of new-born Syrian hamster is deposited in the Russian Collection of Cell Cultures (RCC) in the specialized collection of finite somatic cell cultures of agricultural and game animals at the All-Russian Scientific Research Institute of Experimental Veterinary Medicine n.a. Y.R. Kovalenko (Russian Academy of Agricultural Sciences of farm animals) under No. 89 and intended for reproduction of foot-and-mouth disease virus type A, O, C, Asia-1, CAT-1, CAT-2, CAT-3, and the rabies virus strain "Schyolkovo-51", used for production of antiviral vaccines against foot-and-mouth disease and rabies.
EFFECT: enhanced immunogenicity, infectivity and concentration of cells in industrial suspension cultivation in bioreactors.
SUBSTANCE: claimed invention deals with method of obtaining population of cells, expressing pluripotency markers and markers, characteristic of formed endoderm line. Claimed method includes the following stages (a) obtaining population of cells, expressing markers HNF-3β, GATA-4, Mixl1, CXCR4, and SOX-17, characteristic of formed endoderm line, and (b) cultivation of population of cells under conditions of hypoxia on tissue culture substrate, which is not subjected to preliminary treatment with protein or extracellular matrix, in medium with addition of activin A and ligand Wnt and IGF-1 or insulin, transferrin and selenium, where cultivated cells express markers of formed endoderm line and pluripotency markers.
EFFECT: characterised invention makes it possible to obtain cells, capable of cultivation under conditions of hypoxia, which do not require lines of feeding cells, coatings of complex matrix proteins and preserve differentiation potential.
16 cl, 39 dwg, 9 tbl, 32 ex
SUBSTANCE: group of inventions refers to medicine, namely to immunology, and can be used in laboratory diagnostics, as a test system and a method for measuring interferon alpha (IFN-α) antiviral activity in human blood serum. The test system for measuring IFN-α activity in the human blood serum comprises diploid cells, a virus-containing fluid and standard human interferon-α (IFN-α). As the diploid cells, the test system comprises cells of the characterised line of the diploid cells - human M-20 fibroblasts at 20-33 passages cultured in the medium with added 10% fibrinolytically active plasma (FAP). As the virus, the system contains the A vesicular stomatitis virus (VSV) adapted to M-20 cells, the Indiana strain; the test system additionally contains a viral dye based on two fluorochromes - acriflavine and rhodamine C. The group of inventions also involves a method for measuring IFN-α activity in the human blood serum with the use of the developed system.
EFFECT: using the given inventions enables the quantitative good-reproducible measurement of IFN-α activity in the examined blood serum sampled by the luminescent microscopy of the vital preparations coloured in a fluorochrome-based vital dye.
3 cl, 4 dwg, 1 tbl, 4 ex
SUBSTANCE: invention relates to field of biotechnology, namely to internalisation of therapeutic molecules into cell, and can be applied in medicine. Obtained is composition for delivering molecules of nucleic acids into cells, containing at least one peptide with at least 92% identity to GAAEAAARVYDLGLRRLRQRRRLRRERVRA (SEQ ID NO: 2); IREIMEKFGKQPVSLPARRLKLRGRKRRQR (SEQ ID NO: 3); or YLKVVRKHHRVIAGQFFGHHHTDSFRMLYD (SEQ ID NO: 4), bound to one or several molecules of nucleic acids.
EFFECT: invention makes it possible to increase efficiency of delivery of molecules of nucleic acids into mammalian cell due to peptide, capable of internalisation into mammalian cell with efficiency, constituting at least 200% of efficiency of internalisation of peptide TAT, which has amino acid sequence GRKKRRQRRRPPQ (SEQ ID NO: 1).
8 cl, 16 dwg, 1 tbl, 8 ex
SUBSTANCE: group of inventions relates to methods for the treatment or prevention of diseases, caused by the neovascularisation of the human choroid (neovascular maculopathy), as well to pharmaceutical compositions, containing as an active ingredient at least one peptide from peptides, containing an amino acid sequence, obtained from VEGF-receptor 1 protein, and possessing activity to induce cyctotoxic T-cells in the presence of antigen-presenting cells and at least one peptide from peptides, containing an amino acid sequence, obtained from VEGF-receptor 2 protein and possessing activity to induce cyctotoxic T-cells in the presence of antigen-presenting cells or polynucleotides that code them, where cells of the vascular epithelium, involved into the neovascularisation of the human choroid, express VEGFR-1 receptor protein on the surface of cells.
EFFECT: group of inventions is effective in the treatment or prevention of diseases, caused by the neovascularisation of the human choroid (neovascular maculopathy).
36 cl, 19 dwg, 1 tbl, 1 ex
SUBSTANCE: invention relates to biotechnology. A disclosed polypeptide having the amino acid sequence which has a sequence identity of not less than 80% to the amino acid sequence shown in SEQ ID NO: 1 or 2, revealed in description, which polypeptide has a capable of expressing the polynucleotide activity. Also a polynucleotide encoding D-lactate dehydrogenase originated from DNA construct in which the polynucleotide and a promoter capable of expressing the polynucleotide are linked is introduced. Also described a transformant for production of lactic acid, or transformed yeast, in which the polynucleotide or the DNA construct is introduced. A method of producing D-lactic acid, which comprises the step of culturing the said transformant.
EFFECT: transformant capable of highly producing D-lactic acid compared to the D-lactic acid produced with host cell.
15 cl, 2 dwg, 4 tbl, 13 ex
SUBSTANCE: method comprises immunoaffinity chromatography using mini-antibodies a-hLF-1 and a-hLF-4, which amino acid sequences are presented as SEQ ID NO:1 and SEQ ID NO:2. The invention may be used to obtain highly purified fraction of human lactoferrin.
EFFECT: invention enables to carry out with high efficiency the separation of proteins of lactoferrin of human and goat consisting in milk, using the single-domain mini-antibodies.
6 dwg, 2 ex
SUBSTANCE: invention refers to biotechnology and can be used for recombinant production of human tissue factor (hTF). Constructed is a plasmid pHYP-10ETFCS6 having a length of 5,912 b.p. with a physical map presented on Fig. 2, for expression in a bacterium of the genus Escherichia, which is a precursor of the mutant [C245S] hTF containing an inseparable N-terminal leader peptide containing a deca-histidine cluster and an enterokinase identification sequence fused in a frame with a sequence coding the above mutein fused in the frame with the sequence coding the additional inseparable C-terminal peptide containing the deca-histidine cluster. A method for producing the precursor of the mutein[C245S]hTF contains culturing the producing bacterium in a nutrient medium, recovering inclusion bodies, solubilising the precursor protein, performing a metal chelator chromatography in the denaturation environment, re-folding and diafiltration of the protein solution. A method for producing the mature mutein[C245S] hTF involves detecting the N-terminal leader peptide from the above mutein precursor with using enterokinase and recovering the target protein.
EFFECT: invention enables increasing the level of biosynthesis and yield of pro-coagulation active hTF.
9 cl, 5 dwg, 1 tbl, 7 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to the field of biotechnology, namely to obtaining oligopeptide compounds, containing a motive, interacting with a proliferating cell nuclear antigen (PCNA) and can be used in medicine. The oligopeptide compound consists of 14-70 amino acids and contains. a PCNA-interacting motive, representing [K/R]-[F/Y/W]-[L/I/V/A]-[L/I/V/A]-[K/R], at least one signal sequence of nuclear localisation and at least one signal sequence of penetration into a cell, with the PCNA-interacting motive being located towards an N-end relative to the signal sequence.
EFFECT: invention makes it possible to carry out the efficient treatment of hyperproliferative disorders by the application of the oligopeptide compound in cyctostatic therapy or in radiotherapy as a sensitising substance.
34 cl, 6 dwg, 4 tbl, 8 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: present invention refers to antibodies including human antibodies and their antigen-binding portions, which specifically bind to CCR2, in particular to human CCR2, and can act as CCR2 inhibitors. Anti-CCR2 antibodies are those binding to first and/or second extra-cellular CCR2 loops. The present invention also refers to human anti-CCR2 antibodies and to their antigen-binding portions. The present invention refers to the recovered heavy and light chains of immunoglobulin initiated from human anti-CCR2 antibodies, and to nucleic acid molecules coding such immunoglobulins. The present invention also refers to methods for preparing human anti-CCR2 antibodies and their antigen-binding portions, to compositions containing such antibodies or their antigen-binding portions, and to methods for using antibodies and their antigen-binding portions, and compositions for diagnosing and treating.
EFFECT: invention refers to methods for gene therapy with the use of nucleic acid molecules coding molecules of heavy and light chains of immunoglobulin, wherein the above molecules contain anti-CCR2 antibodies and their antigen-binding portions.
25 cl, 24 dwg, 8 tbl, 17 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology, specifically to a fused protein containing a variant of rodostomin, and can be used in medicine. An ανβ3 integrin selective polypeptide consisting of an amino acid sequence SEQ ID NO:1 conjugated on the N terminal by a linker amino acid sequence containing a combination of the amino acids glycine and serine with a variant of a human serum albumin (HSA) with SEQ ID NO:4.
EFFECT: invention enables the higher therapeutic effectiveness in the diseases related to ανβ3 integrin.
12 cl, 14 dwg, 2 tbl, 7 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology and immunology. Presented are antibodies targeting integrin α2β1 containing humanised anti-integrin alpha-2 (α2) antibodies, as well as a method of treating by the integrin α2 antibodies. The humanised integrin α2 antibodies comprise a variable region of a light chain domain, a constant human light chain domain and a variable constant heavy chain domain of human IgG1, which exhibit the altered effector function. The variable constant heavy chain domain of human IgG1 comprises an S324N substitution. The invention can be used in medicine.
EFFECT: antibodies exhibit complement-dependent cytotoxicity, improved antibody-dependent cell-mediated cytotoxicity and improved CDC and ADCC.
33 cl, 3 dwg, 1 tbl, 2 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology and immunology. What is described is a pharmaceutical composition used for treating and/or preventing pathological bone metabolism and containing this antibody. The invention can be used in medicine.
EFFECT: antibody and its functional fragment specifically recognising human Siglec-15 and possessing the osteoclast inhibitory activity are described.
73 cl, 57 dwg, 4 tbl, 33 ex
SUBSTANCE: present invention relates to biotechnology and provides a α1,6-glucan-containing compound of Helicobacter pylori. The present invention also discloses a conjugate for inducing immune response against H.pylori, which contains said compound conjugated with a carrier protein. The present invention also discloses an immunogenic composition, use of said composition and a method of inducing immune response against H.pylori using said composition. The present invention also discloses immune serum for neutralising H.pylori in mammals, which is obtained by immunising said mammal with an immunogenic composition containing said immunogenic composition. The present invention discloses an antibody which recognises said α1,6-glucan-containing compound of H.pylori, use of said antibody and a method of inducing complement-mediated bacteriolysis of H.pylori strains which express α1,6-glucan using said antibody.
EFFECT: invention improves the effectiveness of immunogenic compositions against Hpylori.
27 cl, 8 dwg, 21 tbl, 11 ex