Thrombin-binding molecules of antibodies and their application
SUBSTANCE: isolated antibody molecule, specifically binding to the thrombin exosite area 1, and an antigen-binding fragment of said antibody, are provided. The use of the antibody molecule in the manufacture of medicaments is considered. A pharmaceutical composition is described as well as a method for treating a thrombin-mediated condition.
EFFECT: use in the treatment of diseases associated with thrombin.
25 cl, 22 dwg, 4 tbl
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to the field of biotechnology, namely, to obtaining inhibitors of adhesion and/or aggregation of platelets, and can be used in medicine. A polypeptide, used as a component of a pharmaceutical composition and in sets for screening of the inhibitors of platelet adhesion or aggregation, is obtained in a recombinant way with the application of a matrix of the salivary gland cDNA of Anopheles stephensi.
EFFECT: invention makes it possible to obtain the polypeptide, possessing inhibiting activity with respect to platelet aggregation and/or inhibiting activity with respect to platelet adhesion.
10 cl, 4 dwg, 5 ex
SUBSTANCE: expression vector is also provided, comprising the polynucleotide and a transformant obtained from S. cerevisiae, for production of the said protein, comprising the polynucleotide or vector.
EFFECT: efficient production of fatty acids with long-chain, having 18 carbon atoms.
9 cl, 3 dwg, 3 tbl
SUBSTANCE: invention relates to biotechnology and genetic engineering. Claimed is expression plasmid vector for monocistronic heterologous expression of recombinant proteins in cells of Chinese hamster ovary (CHO), possessing property of high-frequency integration of expression cassette in said cells, in the following sequence containing region of initiation of plasmid pUC replication with functional gene of ampicilline resistance; site of Epstein-Barr virus terminal repetition (EBVTR), functional promoter of elongation factor 1 alpha gene of Chinese hamster, flanked with 5' untranslated region of said gene; site for cloning open reading frame of recombinant protein (polylinker); functional terminator of elongation factor 1 alpha gene of Chinese hamster, flanked with 3' untranslated region of said gene, promoter, functioning in mammalian cells; gene of antibiotic resistance; and functional terminator of antibiotic resistance gene. Also claimed is line of cells - recombinant protein producers and method of obtaining recombinant protein with application of said cells.
EFFECT: invention makes it possible to increase stability and productivity of expression systems of recombinant proteins.
13 cl, 7 dwg, 6 tbl, 8 ex
SUBSTANCE: invention refers to biotechnology. What is presented is nucleic acid coding protein possessing acetyl-CoA-carboxylase activity making up the deficiency of acetyl-CoA-carboxylase in yeast, wherein a nucleotide sequence is specified in nucleic acid, which contains a nucleotide sequence: (a) coding protein consisting of the amino acid sequence SEQ ID NO:2; (b) hybridised in the hard conditions with nucleic acid complementary to SEQ ID NO:1; (c) SEQ ID NO:1; and (d) hybridised in the hard conditions with nucleic acid consisting of the complementary nucleic sequence coding protein SEQ ID NO:2; wherein SEQ ID NO:1 and 2 are disclosed in the description. There are also described: acetyl-CoA-carboxylase (SEQ ID NO:2) increasing the host-specific arachidonic acid content; a recombinant vector containing the above nucleic acid; and a cell transformed by the above vector for producing the fatty acid composition rich in arachidonic acid. What is presented is a method for producing the fatty acid composition involving culturing the above cell and collecting the fatty acid composition from the transformed cell culture.
EFFECT: invention enables producing the fatty acid composition rich in arachidonic acid in the host cell.
11 cl, 8 dwg, 5 tbl, 8 ex
SUBSTANCE: claimed inventions deal with a modified protein, nucleic acid, coding such protein, a vector, containing nucleic acid, and a carrier for biotin binding, which such protein is immobilised on. The characterised modified biotin-binding protein is obtained by the introduction of a mutation from one to several amino acid residues into a sequence, represented in SEQ ID NO:2, or an amino acid sequence, identical to the said sequence by 98% or more, and the presence of the biotin-binding activity, where at least one residue, selected from the group, consisting of residues from 1) to 4), presented below, is substituted with the residue of acidic amino acid or residue of neutral amino acid; 1) residue of arginine in position 104 SEQ ID NO: 2; 2) residue of lysine in position 141 SEQ ID NO: 2; 3) residue of lysine in position 26 SEQ ID NO: 2 and 4) residue of lysine in position 73 SEQ ID NO: 2.
EFFECT: claimed inventions make it possible to obtain the biotin-binding protein and can be applied for biotin binding.
14 cl, 6 dwg, 11 tbl, 3 ex
SUBSTANCE: invention relates to biotechnology and gene engineering. A method for selecting at least one transfected eukaryotic host cell expressing a target product, the eukaryotic host cells comprise at least an introduced polynucleotide encoding the target product, an introduced polynucleotide encoding a DHFR enzyme using at least one expression vector, providing a plurality of eukaryotic host cells, whose viability is dependent upon folate uptake, wherein the said host cells comprise at least a foreign polynucleotide encoding the target product, a foreign polynucleotide encoding a DHFR enzyme, culturing the said plurality of the eukaryotic host cells in a selective culture medium comprising folic acid in a concentration of 12.5-50 nM combined with a concentration of MTX of 2.3-500 nM, selecting at least one eukaryotic host cell expressing the target product.
EFFECT: described is a method of the target product and culture medium preparation.
11 cl, 2 tbl, 2 ex
SUBSTANCE: invention relates to field of medicine, genetic engineering and biotechnology. Claimed is method of providing tumour-free tissue-substituting therapy on the basis of obtained from adult somatic cells induced pluripotent stem cells (iPSC), where the latter are genetically modified by means of artificial chromosome (AC), carrying bicistronic cassette with suicide-gene and gene of sensitivity to antibiotic under control of regulatory element, specific for pluripotent stem cells, with modified iPSC being selected in presence of respective antibiotic, absence of AC integration into iPSC genome is confirmed, are introduced into recipient organism directly, without preliminarily differentiation in vitro, after 1-14 days patients are given a 5-10-day course of therapy with inductor of toxicity of suicide-gene product.
EFFECT: invention makes it possible to reduce to zero risk of cancer transformation of transplanted cells and development of teratomas, increase clinical efficiency of recovery of cell mass of injured organ or tissue, reduce waiting time for potential recipients.
6 cl, 1 tbl, 1 dwg
SUBSTANCE: synthetic DNA is proposed, encoding human erythropoietin, having the sequence Seq ID No. 1, comprising its expression vector, the method of production of erythropoietin producer strain, and a strain of a Chinese hamster ovary cells - producer of recombinant human erythropoietin, deposited under the number RKKK(P) 761 D.
EFFECT: invention enables to increase the expression level of recombinant human erythropoietin.
5 cl, 1 tbl, 8 dwg, 4 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to molecular biology and medicine. There are presented ribonucleic acids of general formula (NuGlXmGnNv)a and derivatives thereof as an immunostimulating agent, compositions containing them.
EFFECT: treating cancer diseases, infectious diseases, allergic and autoimmune diseases.
21 cl, 2 dwg, 5 tbl
SUBSTANCE: group of inventions relates to field of biotechnology. Method of selecting food diet includes determination of food diet or substance, which increase amount of microRNA, present in mammalian milk, applying correlation of miroRNA profiles in milk and food diet, obtained by mammal, or substance, contained in food diet, as index. Profiles of microRNA in milk, identified before and after dietary intake, are compared, and if amount of at least one type of microRNA, identified after intake, is higher than that identified before intake, it is supposed that diet increased amount of microRNA in milk. Method additionally includes measurement of microRNA in milk and microRNA profiles in serum of plasma, and if amount of microRNA, contained both in milk and serum or plasma, after dietary intake increases in milk by 1.47 times or higher in comparison with that identified in serum or plasma, it is supposed that diet increases amount of microRNA in milk.
EFFECT: application of invention group makes it possible to obtain breast milk, possessing immunostimulating action.
6 cl, 7 dwg, 11 tbl, 5 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention deals with application of composition, which includes hydrolysate of pea protein and/or peptide for obtaining composition for treatment and/or prevention of infection Helicobacter pylori (versions), as well as such compositions (versions). Characterised compositions include lipid, protein and carbohydrate component, in which protein component includes protein source, which consists of pea protein hydrolysate, obtained by hydrolysis with protease, different from chymotrypsin, or from peptides, selected from group, which consists of Xaan-Asp-Phe-Leu-Glu-Asp-Ala-Phe-Asn-Val-Asn-Arg-Xaam and Xaan-Glu-Leu-Ala-Phe-Pro-Gly-Ser-Ala-Gln-Glu-Val-Asp-Arg-Xaam, where each Xaa independently can be any amino acid, and n and m are integer numbers, independently varying from 0-10, where peptide is contained in pea protein hydrolysate, or from both.
EFFECT: claimed inventions make it possible to treat or prevent diseases, caused by Helicobacter pylori infection, and/or diseases, associated with infection Helicobacter pylori in mammals.
17 cl, 2 tbl, 1 ex
FIELD: food industry.
SUBSTANCE: invention relates to a method for manufacture of multiple products from aquatic plant species biomass. Biomass is obtained, destroyed and separated to produce juice and solid phase; the juice is filtered and clarified. Protein is coagulated from the clarified juice to produce broth including a wet protein concentrate. The said concentrate is separated from broth. The wet protein concentrate is dried to obtain dry protein concentrate. The solid phase is used for wet biological raw materials production. The said biological raw materials are dried to produce at least one product selected from among dry biological raw materials and meal rich in carbohydrates. At least 50% of protein in the multiple products is present in dry protein concentration.
EFFECT: method is environmentally friendly and allows production of multiple products selected from among dry biological raw materials and meal rich in carbohydrates.
35 cl, 39 dwg, 7 tbl, 25 ex
SUBSTANCE: invention relates to biotechnology of obtaining haemostatic medications. Claimed is method of separating purified fibrinogen concentrate, free of viruses and ballast proteins. Solubilisation of cryoprecipitate of fresh frozen human plasma is realised. Fibrinogen is precipitated with 20-30% PEG solution. Separated sediment is dissolved in buffer with sodium citrate and sodium chloride. Virus inactivation of solution by solvent-detergent method is carried out in presence of 1-3% Tween-80 and 0.1-1.5% of tri-n-butylphoshate. Obtained concentrate is purified from products of virus inactivation and solvent-detergents by triple extraction with liquid paraffin. After that, obtained fibrinogen concentrate is re-precipitated with 1.0-2.5 M glycine solution. Sterile filtration and lyophilic drying with further corking of lyophilisate under vacuum and thermal inactivation are performed.
EFFECT: invention makes it possible to obtained lyophilised form of fibrinogen concentrate with approximately 55% output.
SUBSTANCE: method comprises the steps of destroying the bodies of inclusion, renaturation and purification of protein. Before renaturation, the preliminary purification of protein is carried out using chromatography on Q-Sepharose and SP-Sepharose using the combined columns with Q- and SP-Sepharose. After renaturation chelate and ion-exchange chromatography of recombinant prourokinase M5 is carried out without intermediate elution of the target protein with use of metal-chelate sorbent activated with ions Co2+ or Zn2+.
EFFECT: invention enables to select prourokinase M5 from inclusion bodies, comprising the present prourokinase, with high yield.
4 cl, 2 tbl, 4 ex
SUBSTANCE: invention relates to the field of obtaining and separation of single-domain molecules (SDAB). Described is a method of the separation or purification of the SDAB molecule, which represents a trivalent molecule of a ATN-103 nanobody, targeting TNFα and HAS, from a mixture, containing the said SDAB molecule and one or more polluting substances. The mixture is brought in contact with a cation-exchange carrier under conditions, which make it possible for the SDAB molecule to bind with the carrier or be absorbed on the carrier. One or more polluting substances are removed and SDAB is selectively eluted from the carrier. The conductivity of a conditioning medium (CM), used for the carrier loading, constitutes from approximately 12 to 9 mS/cm and pH under conditions of loading is corrected to a value from 4.0 to 4.3. The buffer for elution corresponds to approximately 50 mM of sodium chloride or less and has pH from approximately 5.5 to 7.2. Disclosed is a method or a process of obtaining recombinant SDAB of ATN-103. A host-cell is supported in the conditions at which recombinant ATN-103 SDAB is expressed. The mixture of molecule SDAB and one or more polluting substances is obtained. ATN-103 SDAB is purified or separated with the application of cation-exchange chromatography, as said above.
EFFECT: application of the invention provides new methods of the separation or purification of the nanobody, which can be applied in obtaining the ATN-103 nanobody.
19 cl, 4 dwg, 6 ex
SUBSTANCE: method of obtaining of a complex of antimicrobic peptides of an insect includes infecting of adipose body of an insect at a larval instar with Micrococcus luteus A270 and Escherichia coli D31 bacteria with the subsequent extraction of adipose body of an insect at a larval instar. The adipose body of an insect is placed into a nutrient medium containing water solution of sugars, inorganic salts and the antibiotic meropenem in pre-set ratio and incubated during a day with the subsequent elution of the complex of antimicrobic peptides of an insect from cultural liquid by the method of reverse-phase chromatography on the column Vydac C18 at the linear gradient of acetonitrile from 0% up to 50%.
EFFECT: invention allows to simplify a method of obtaining antimicrobic peptides.
5 dwg, 4 ex
SUBSTANCE: invention refers to peptide chemistry and concerns producing tripeptide diacetate H-β-Ala-Pro-DabNHBzl referred to a biologically active compound used in cosmetic industry as an active component for cosmetic products, particularly for stimulating skin rejuvenation, tightening and prevention. The method is based on the 6-staged synthesis and is free from the stages of setting and releasing the protective groups. The method involves proline β-chlorpionyl chloride acylation followed by producing N-(3-chlorpropionyl)proline pentafluorphenyl ester in the presence of N,N'-dicyclohexyl carbodiimide. The above pentafluorphenyl ester is condensed thereafter with glumatic acid monomethyl ester to produce N-(β-chlorpropionyl)-Pro-Glu(δ-OMe)OH. That is followed by benzylamine amidation in a combination with ammonolysis and chlorine substitution by an amino group. That enables producing the tripeptide β-Ala-Pro-Glu(δ-NH2)NHBzl; Hofmann rearrangement is conducted with the use of iodo-benzene diacetate to produce a target product.
EFFECT: method is characterised by simplicity, effectiveness; it is cost-effective and uses more accessible and cheap agents.
SUBSTANCE: method of obtaining SSI comprises the following steps. The strain Yersinis pestis KM 1279 is grown on 1.5% agar LB, the bacteria are washed three times with cold buffered normal saline. The bacteria are pelleted by centrifugation, suspended in a solution of 5 mM NaOH, kept at 37°C for two hours and the cells are pelleted by centrifugation. The supernatant is selected and the procedure as repeated three times, three supernatants are combined and filtered through the nitrocellulose membrane. The filtrate is extracted three times with the mixture of chloroform-methanol-water in a ratio of 5:2:1. The chloroform fractions are separated by centrifugation, combined and freed from water-soluble impurities. The aqueous fraction is separated by centrifugation and removed, and the chloroform fraction is dried in a vacuum rotary evaporator and the dry preparation SSI is obtained. The proposed SSI is characterised with brown colouring of dry crystals, hydrophobic properties, fluorescence in ultraviolet, lipopeptide nature, the presence of iron ions, the molecular weight of 380.6 Da.
EFFECT: inventions enable to obtain the natural regulator of virulence of plague agent.
2 cl, 6 dwg, 6 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology. What is presented is a method for preparing a recombinant protein of type III interferon-like factor (ILF III) of the producing strain E. coli. The inclusion bodies E. coli are washed and dissolved with using 2% aqueous γ-cyclodextrin. That is followed by the sequential Ni-Sepharose, Q-Sepharose and SP-Sepharose chromatographic procedures. Refolding of a target protein is performed with using a mixture of cysteamine and cystamine at pH 10.5. The Amberchrome Profile XT20, Amberchrome Profile HPR10 and Kromasil 300-5C18 chromatographic procedures are sequentially performed.
EFFECT: invention enables optimising the ILF III purification environment at the stage of washing and dissolving the inclusion bodies Ecoli and provides 12% target protein yield.
3 dwg, 4 ex
SUBSTANCE: invention relates to biotechnology, specifically to immunostimulating compounds and may be used in medicine. An immunostimulating peptide of an amino acid sequence XLYDKGYTSKEQKDCVGI, where N-terminal X is N-acetylalanine, and may be covalently linked to fatty acids, selected from C2-C25, to form PDAG (peptidyl-2,3-diacylglycerides). The resulted compound may be included in pharmaceutical formulations for stimulating an immune response.
EFFECT: invention provides efficient stimulation of an immune response in subjects and may enhance the immunogenicity of the antigenic peptide when administered with PDAG.
29 cl, 10 dwg, 9 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to field of immunology and biotechnology. Claimed is monoclonal antibody or its functional fragment, where said antibody and fragment bind with activated protein C and inhibit anticoagulant activity, but do not bind and do not inhibit activation of inactivated protein C, where said antibody is obtained by immunisation of mammal by APC and screening of binding ability of said antibody with APC, but not with protein C. Also described is pharmaceutical composition for treating diseases associated with anticoagulation activity of APC, including said antibody in effective amount and pharmaceutically acceptable carrier. Claimed are: method of inhibiting anticoagulation activity of activated protein C in subject, method of inhibiting amidolytic activity of activated protein C in subject, method of treating subject, requiring blood coagulation; method of treating subject with haemophilia; method of modulating haemostasis in subject; as well as method of modulating thrombogenesis in subject, which include introduction of effective quality of said antibody to subject. In addition, described is method of treating subject with sepsis, including introduction of effective quality of said antibody and activated protein C.
EFFECT: invention makes it possible to obtain monoclonal antibody or its functional fragment, where said antibody and fragment bind with activated protein C and inhibit anticoagulation activity, but do not bind and do not inhibit activation of inactivated protein C.
17 cl, 11 dwg, 6 ex