Structures of fusion protein in spider silk for binding to organic target
SUBSTANCE: invention relates to obtaining a polymeric protein structure capable of selective interaction with organic targets and can be used in medicine. The polymer protein structure contains a recombinant fusion protein, as a recurring structural element, which includes parts B, REP and CT.
EFFECT: invention allows for producing a protein structure, which can be used as an affine medium and scaffold material for cells.
41 cl, 34 dwg, 5 tbl, 28 ex
FIELD: food industry.
SUBSTANCE: protein produced of milk is plasticised together with a plasticiser at a temperature of 60°C - 140°C under mechanical stress conditions; then fibres are moulded by means of a draw die. The plasticiser is chosen from the group including a polysaccharide water solution, glycerine, ethylene glycol or these substances mixture.
EFFECT: produce fibres have significant tensile strength.
13 cl, 1 tbl, 3 dwg, 1 ex
SUBSTANCE: composition of forming solution for formation of biopolymeric fibres includes polyethylene oxide, chitosan, organic acid and water. Or composition of forming solution for formation of biopolymeric fibres includes chitosan, water and component, selected from polyethylene oxide, polyvinyl alcohol or polyvinylpyrrolidone. Chitosan with molecular weight 30-40 kDa and polyethylene oxide with molecular weight 2000-8000 kDa are used. Composition additionally contains cellulose diacetate and biologically active substances. Method of forming solution preparation includes mixing of components in powder-like state and their dissolution with mixing to homogenous state. Linen of biomedical purpose, formed from biopolymer chitosan-based fibres, in order to obtain from it bandage for wound treatment, is subjected to swelling in physiological solution or distilled water. Additionally linen of biomedical purpose is subjected to thermal processing, after which it is placed into distilled water or physiological solution for swelling. Or linen is additionally processed with alkaline reagent, with further washing with distilled water, after which it is placed into distilled water for swelling. For wound treatment linen of biomedical purpose is applied on wound in form of biological bandage.
EFFECT: invention makes it possible to obtain biopolymer fibre with application of optimal compositions of textile composition based on chitosan and non-toxic polymer for stable electroformation of defect-free fibres, methods of modification of linen of biomedical purpose for obtaining bandage in form of wound covering are ecologically clean and economical.
2 tbl, 73 ex, 5 dwg
FIELD: textile industry.
SUBSTANCE: invention relates to fabrication of artificial fibers possessing antibacterial activity, in particular to fabrication of chitosan-containing fibers. Process comprises preparation of solution of chitosan in acetic acid, to which concentrated sodium hydroxide solution is added in order to perform mercerization. Mercerized chitosan is then subjected to xanthogenation to produce chitosan viscose. The latter is molded, finished, and dried. According to second option, thus obtained chitosan viscose is blended with cellulose viscose followed by molding, finishing, and drying resulting chitosan-containing blend. According to third option, chitosan-containing alkali solution is supplemented by pulped cellulose for subsequent joint mercerization, xanthogenation, finishing, and drying.
EFFECT: imparted antibacterial activity.
3 cl, 1 tbl, 3 ex
FIELD: process of production of synthesized textile fiber containing phytoprotein, applicable in the textile industry.
SUBSTANCE: the fiber contains exactly or less than 94 parts of the total percentage of materials of polyvinyl alcohol and exactly or more than 21 parts of phytoprotein. The method for fiber production consists in production of a spinning solution of polyvinyl alcohol containing phytoprotein, precipitation in a coagulation bath, drafting, thermofixation, finishing and acetalization.
EFFECT: produced phytoprotein synthesized fiber with a maximum air permeability and possessing properties similar to those of cashmere, reduced duration of the production cycle and enhanced output.
18 cl, 13 ex
FIELD: artificial fibers manufacturing, in particular chitosan-containing fiber with antifungal and antibacterial activity.
SUBSTANCE: chitosan is dissolved in acetic acid solution and passed through sputtering dryer. Then obtained activated chitosan with porous structure in form of microparticles is brought into reactions of mercerization, xantation and wet extrusion.
EFFECT: chitosan-containing fiber with antibacterial activity and good biodegradability.
4 ex, 1 tbl
FIELD: artificial fibers manufacturing, in particular chitosan-containing fiber with antifungal and antibacterial activity, useful in medicine, industry, public health.
SUBSTANCE: claimed method includes chitin crushing, its single treatment with sodium hydroxide solution up to de acetylation coefficient of 91 % or more followed by treatment with carbon disulfide and extrusion from obtained solution. Fiber obtained from chitosan viscose has ratio of chitosan:chitin more than 10:1.
EFFECT: industrial method for production of chitosan-containing fiber with biocompatibility, antibacterial activity, good biodegradability.
2 cl, 5 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 the field of biotechnology and can be applied for the selection of affine molecules. Constructed is a plasmid DNA pGST/APT/X 6193 bp long, with the weight of 4.09 MDa, containing as a genetic marker the gene β-lactase and unique sites of the recognition of restriction endonucleases, located at the following distance to the right from the site NdeI: BgIII 665bp, BamHI 779 bp., XhoI 801 bp. The plasmid pGST/APT/X consists of a NdeI-XhoI DNA fragment of the commercial plasmid pET22b(+) and nucleotide sequence SEQ ID NO:1, inserted by sites NdeI-XhoI and combining in its composition the sequences, which code a fragment of a gene of glutathione-S-transferase, a peptide substrate of a lethal factor of anthrax RRKKVYPYPME, peptide GGLNDIFEAQKIEWHED, biotinylated in vivo under the influence of E.coli biotin-ligase, and a linker sequence, substituted by genetic-engineering manipulations by sites of restriction endonucleases BamHI and XhoI for a sequence, coding a target protein B-subunit of shiga-toxin I.
EFFECT: invention makes it possible to carry out the selection of highly specific aptamers to the said target protein.
2 cl, 4 dwg, 4 ex
SUBSTANCE: invention relates to biotechnology, specifically to novel hetero-multimeric proteins obtained from modified ubiquitin, and can be used in medicine to treat or diagnose diseases associated with hyperprodution of the extradomain B of fibronectin (ED-B). The protein includes two monomeric ubiquitin links which are differently modified through substitutions of at least 6 amino acids in positions 4, 6, 8, 62, 63, 64, 65 and 66 of SEQ ID NO: 1. In the first monomer link the substitutions include: F4W, K6(H, W or F), Q62N, E64(K, R or H), S65(L, F or W), T66(S or P), and in the second monomer link: K6(T, N, S or Q), L8(Q, T, N or S), Q62(W or F), K63(S, T, N or Q), E64(N, S, T or Q), S65(F or W), T66(E or D).
EFFECT: invention enables to obtain a modified heterodimeric ubiquitin protein, capable of binding with ED-B with high affinity.
28 cl, 18 dwg, 3 tbl, 7 ex
SUBSTANCE: invention relates to the field of biotechnology and can be used for the identification of heteromultimeric ubiquitins possessing an ability to bind with a ligand-antigen. The method includes the contact of a totality of heterodimeric modified ubiquitins, including two ubiquitin monomers, bound to each other by a head-to-tail scheme, with the potential ligand in a display way. Each of the said monomers is modified in a different way and contains 5-8 substitutions in positions 2, 4, 6, 8, 62, 63, 64, 65, 66 and 68 SEQ ID NO:1. After that, a heterodimeric modified protein, which has bound with the ligand with the binding affinity Kd in the range of 10-7-10-12 M and the monovalent binding activity. Claimed are DNA-libraries, responsible for obtaining a population of the said heteromultimeric ubiquitins, as well as libraries of proteins, obtained by the expression of the said DNA-libraries.
EFFECT: invention makes it possible to obtain the novel bonding proteins based on heteromultimeric ubiquitin, capable of specific high affinity binding with the selected ligands.
8 cl, 17 dwg, 4 ex
SUBSTANCE: invention relates to the field of biotechnology, in particular to the lentiviral delivery of apoptin into tumour cells, and can be used in medicine. The method includes obtaining a lentiviral construct, expressing modified apoptin, fused with a sectretory signal of lactotransferrin and a transduction signal (ST-CTP-apoptin), with the further obtaining of recombinant lentiviral particles, defective by replication and carrying the modified apoptin, which are later introduced into T-lymphocytes (TILs), obtained in the surgical ablation of the tumour or in the process of obtaining a biopsy, possessing the ability to penetrate into tumour cells. After that, obtained TILs are autotransplanted to the said patient.
EFFECT: invention makes it possible to increase the ability of apoptin to penetrate into tumour cells and produce an oncolytic effect with respect to all the tumour cells.
2 dwg, 3 ex
SUBSTANCE: group of inventions refers to biotechnology. What is presented is a precursor protein for the recombinant preparation of peptides having a length of a sequence of 5 to 70 amino acid residues. The above protein contains a splittable recurrent sequence of target peptide elements (Pep) and auxiliary peptide elements (Aux) of general formula: (Pep-Aux)x or (Aux-Pep)x, wherein x>1. The elements Aux contain a self-assembled peptide element (SA). The elements Pep contain an amino acid sequence having a length of the sequence of 5 to 70 amino acid residues; ends of the elements comprise splittable sequences, which enable releasing the elements Pep from the precursor protein by specific splitting. There are also presented a nucleic acid molecule with a nucleic acid sequence coding the above precursor protein, an expression cartridge and a recombinant expression vector containing the sequence of the above nucleic acid. There are also presented a variant of the precursor protein, a method for producing the target peptide Pep, as well as using an amphiphilic peptide for the recombinant production of the antimicrobial target peptide.
EFFECT: group of inventions provides the higher yield of the target peptides.
24 cl, 11 dwg, 1 tbl, 7 ex