Derivatives of blood coagulation factors vii and viia, conjugates and complexes containing them and their application
SUBSTANCE: invention relates to production of blood coagulation derivatives VII and VIIa, their conjugates with polymers capable of increasing the half-life of blood from the bloodstream, complexes containing them obtained by carrier binding to the conjugate, to genes encoding the derivatives, to expression vectors containing these genes, transformants with introduced expression vectors, to methods for their preparation, pharmaceutical compositions and treatment methods, and can be used in medicine to prevent or treat hemophilia or to improve blood coagulation. A FVII derivative is obtained which is linked at its C-terminus to the peptide linker for binding to a non-peptidyl polymer capable of increasing the half-life of FVII or FVIIa from the blood stream. At that, the peptide linker is a partial sequence of superoxide dismutase SOD1, its mutated sequence or GGGGSC sequence.
EFFECT: invention allows to obtain an FVII derivative capable of binding to a carrier increasing the half-life from the blood stream while maintaining FVII activity.
42 cl, 8 dwg, 3 tbl, 8 ex
SUBSTANCE: invention relates to biochemistry, particularly to an isolated nucleic acid molecule which codes a polypeptide, having delta-9-elongase activity, as well as a purified polypeptide having delta-9-elongase activity, which codes said isolated nucleic acid molecule. The invention also discloses an expression vector containing said isolated nucleic acid molecule, as well as a host cell, a transgenic plant and a transgenic seed containing same.
EFFECT: invention provides efficient production of polyunsaturated fatty acid-enriched oils.
18 cl, 9 dwg, 9 tbl, 6 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: present invention refers to biotechnology and represents a purified recombinant urate oxidase (uricase) characterised by the tetramer and octamer content of 95% and more of the total molecule count. The invention also discloses a purified preparation of the above urate oxidase possessing lower immunogenicity, a conjugate of the above urate oxidase and polyethylene glycol or polyethylene oxide used to reduce mammalian tissue or liquid uric acid, purified fragments of the above urate oxidase, a pharmaceutical composition containing the above urate oxidase and a method for purifying the above urate oxidase with maintaining its uricolytic activity, involving the separation and removal of uricase aggregates bigger than octamers.
EFFECT: present invention enables producing the preparations of urate oxidase of lower immunogenicity.
23 cl, 6 dwg, 6 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 a novel method of obtaining fluorescent catecholamines selected from dopamine and adrenalin, and their metabolites, selected from homovanillic and vanillin-mandelic acids, by a method of derivation. The compounds can be used as highly sensitive and selective markers for the determination of various diseases. The method of derivation includes oxidation of the initial compounds and their interaction with amines that form condensed structures in a medium of the CAPS-buffer solution or glycin - KOH 0.2 mM hydrogen peroxide in the presence of horseradish peroxidase as a catalyst. The process in preferably carried out in a 0.1 M buffer solution with the concentration of horseradish peroxidase 0.01-1 mcM; concentration of hydrogen peroxide - 100 mcM, amine concentration - 0.1-33 mM; concentration of catecholamines and metabolites - 0.03-1 mcM.
EFFECT: method is simple and producible as it does not require higher temperature and is realised in a water solution.
2 cl, 2 dwg, 3 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions relates to biotechnology. Claimed is cell of mycelial fungus Penicillium canescens with removed catabolite repression and arabinose induction, producing xylanase and laccase. Cell is transformed with plasmid, containing promoter of gene bgaS, bgaS leader peptide, xylanase-coding DNA fragment, bgaS terminator, β-lactamase gene and pMB1 replicon, and plasmid, which contains promoter of gene bgaS, bgaS leader peptide, laccase-coding DNA fragment, bgaS terminator, β-lactamase gene and pMB1 replicon. Also claimed is method of enzyme preparation of xylanase and laccase with application of said cell of Penicillium canescens.
EFFECT: group of inventions provides increased output of target enzymes.
10 cl, 9 dwg, 3 tbl, 4 ex
SUBSTANCE: invention relates to the use of the concentrate of the culture liquid of the strain Trichoderma harzianum Rifai, deposited in the Russian National Collection of Industrial Microorganisms under the number of RNCIM F-180 as an inhibitor of Andis virus of potato mottling.
EFFECT: invention enables to reduce losses of potato from the plant infection with the Andis mottling virus.
SUBSTANCE: group of inventions relates to biotechnology and can be applied in creation of analytic methods with application of peroxidases. Method includes preparation of substrate mixture, introduction of peroxidases in substrate mixture with the following registration of intensity of formed luminescence. Substrate mixture includes the following components, given in final concentrations: buffer solution 10-125 mM, luminol 0.05-8 mM, hydrogen peroxide 0.05-8 mM, 4-aminopyridines 0.1-10 mM, N-carboxyphenothiazine, or N-(carboxymethyl)phenothiazine, or N-(2-carboxyethyl)phenothiazine 0.1-10 mM. And pH of buffer solution constitutes 7.9-9.0.
EFFECT: invention ensures obtaining of high intensity of chemiluminescence and, accordingly, high sensitivity of peroxidase activity determination.
4 cl, 8 ex
SUBSTANCE: invention relates to biotechnology and discloses isolated polynucleotides which encode Δ8 desaturase. The invention also relates to Δ8 desaturases themselves, which encode isolated polynucleotides, expression vectors containing isolated polynucleotides, host cells containing expression vectors and methods of producing Δ8 desaturase and polyunsaturated fatty acids selected from a group consisting of dihomo-gamma-linolenic acid (DGLA), ω3-eicosatetraenoic acid (ω3-ETA) and any combinations thereof.
EFFECT: invention widens the range of Δ8 desaturases used to produce polyunsaturated fatty acids.
12 cl, 12 dwg, 14 tbl, 6 ex
SUBSTANCE: invention relates to field of biochemistry, in particular to genetic construction for expression of threonine-insensitive aspartate kinase in plant, where construction contains promoter, specific for phase of aging, functionally connected with coding sequence, which codes polypeptide, possessing activity threonine-insensitive aspartate kinase, and in which specific to aging promoter is selected from group, consisting of SAG12, SAG13, SAG101, SAG21 and SAG18, or their functional variants or fragments. Described are vector, plant cell, transgenic plant, material for plant reproduction, collected leaves, smoking tobacco, which contain claimed genetic construction. Also described is method of obtaining transgenic plant, which possesses ability to accumulate threonine in fading leaves in larger amount than in leaves of corresponding plant of wild type, and method of increasing the level of threonine in leaves of aging plant to the level of threonine, higher than the content of threonine in leaves of plant of wild type without impairing plant adaptation with application of said genetic construction.
EFFECT: invention makes it possible to obtain transgenic plant, which possesses ability to accumulate threonine in fading leaves in larger amount than in leaves of plant of wild type.
22 cl, 16 dwg, 2 tbl, 4 ex
SUBSTANCE: invention is the truncated mutant luciferase MLM4 of Metridia longa with the size of -16 kDa with improved properties for use as a genetically encoded bioluminescent reporter for visualisation of molecular processes in living cells. The amino acid substitutions I69L, N74E, K125V, W139F are introduced in the sequence of the truncated luciferase MLM4 of Metridia longa with the size of -16 kDa.
EFFECT: invention enables to provide the luciferase with improved thermal stability and the spectrum shift to long-wave region, which determines an overall increase in sensitivity of the bioluminescent reporter when used in living cells and organisms.
4 dwg, 1 tbl, 3 ex
SUBSTANCE: invention relates to field of biochemistry, in particular to antibody or its antigen-binding fragment, which is specifically bind with human TNFα. Also disclosed are: separated molecule of nucleic acid, which codes said antibody, vector of expression, which contains said molecule of nucleic acid, and host cell, which contains said vector, for expression of said antibody. Disclosed are pharmaceutical composition for treatment of TNFα-mediated disease, which contains therapeutically effective quantity of said antibody, and method of treating TNFα-mediated disease with application of claimed composition.
EFFECT: invention makes it possible to effectively treat TNFα-mediated diseases.
16 cl, 9 dwg, 2 tbl, 2 ex
SUBSTANCE: invention relates to field of biochemistry, in particular to isolated version of serine protease, containing, at least, from five to eight amino acid substitutions in positions, selected from the group, consisting of positions12, 14, 15, 16, 24, 35, 36, 39, 49, 54, 61, 63, 64, 65, 67, 69, 75, 76, 78, 79, 81, 86, 92, 93, 99, 109, 112, 121, 123, 125, 127, 143, 159, 179, 181, 184, 189, where substituting amino acid is selected from the group, consisting of F, G, L, V, I, S, A, Y, K, W, M, P, N, Q, T, E, H, R, C, N and D, where said substitutions are made in positions, equivalent to positions in protease Cellulomonas 69 B4, as well as to molecule of nucleic acid, which codes it. Described are: expression vector, which contains said molecule of nucleic acid, as well as host-cell, containing it. Also disclosed are cleaning composition, animal food, composition for fabric of leather processing, which contain said version of serine protease, as well as cleaning method with application of said cleaning composition.
EFFECT: claimed invention has improved caseinolytic activity and/or thermal stability, and/or stability in LAS, and/or improved casein hydrolysis in comparison with protease, which does not have said number of substitutions.
63 cl, 7 dwg, 21 tbl, 21 ex
SUBSTANCE: claimed invention relates to the field of biotechnology. Claimed are versions of a humanised anti-CD79b antibody, each of which is characterised by the presence of a light and heavy chain and a set of 6 CDR with a determined amino acid sequence. An epitope of the antibody from 11 amino acids is determined by the Biacore method. Disclosed are: an immunoconjugate of the antibody with a medication or means for inhibiting cell growth, where the antibody is bound with means covalently, and versions of the composition, based on an effective quantity of the immunoconjugate or the antibody, used for inhibiting B-cell proliferation; as well as a method of determining CD79b in a sample with the application of the antibody. Described are: an antibody-coding polynucleotide, as well as an expression vector and an isolated cell, containing the vector for obtaining the antibody. Disclosed are versions of applying the antibody or immunoconjugate for obtaining the medication for inhibiting the growth of CD79b-expressing cells for the treatment of an individual, affected with cancer, for the treatment of proliferative disease or for inhibiting B-cell proliferation.
EFFECT: invention provides novel antibodies, which can find further application in the therapy of proliferative CD79b-associated diseases.
91 cl, 8 tbl, 9 ex, 20 dwg
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: invention relates to genetic engineering and biotechnology. Disclosed is a method of evaluating bioactivity of chemical compounds, where the first step includes transient transfection of cell line HEK 293 with plasmid vector pX-Y-neo (X is any eukaryote transcription factor, Y is a proteotypic peptide corresponding to said transcription factor), which contains a minimal human adenovirus type 5 promoter; a green fluorescent protein gene; a nucleotide sequence which codes the binding site of the transcription factor; a nucleotide sequence which codes the proteotypic peptide; a neomycin resistance gene; the second step includes determining the activity of the transcription factor via fluorescent analysis and chromatographic-mass spectrometer measurement of the content of the proteotypic peptide in the transfected cell culture in the presence of the test substance compared to a transfected intact cell culture.
EFFECT: invention provides fast and highly sensitive evaluation of bioactivity of chemical compounds.
2 dwg, 1 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
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to the field of biotechnology, namely to obtaining insulin analogues, and can be used in medicine as a medication for reducing the glucose level in a patient's blood. An insulin analogue contains a polypeptide B-chain, including halogenated phenylalanine in B24 position, which provides an increased stability in comparison with non-halogenated insulin or insulin analogue. Halogenated phenylalanine represents ortho-monofluorophenylalanine, ortho-monobromophenylalanine or ortho-monochlorophenylalanine.
EFFECT: halogenation-conditioned insulin stabilisation makes it possible to simplify the treatment of patients with diabetes mellitus in developing countries, where there is no access to refrigerating equipment.
11 cl, 8 dwg, 7 tbl
SUBSTANCE: invention relates to biotechnology and fundamental virology. Method includes obtaining chimeric virus by inserting gene of protein of icosahedral low-copy phloem-restricted virus (ILCPRV) envelope into effective viral vector based on tobamovirus RNA. After that plant is infected with obtained chimeric virus for it to multiply and accumulate in plant tissues. Finally, separation of chimeric virus from plant tissues is performed. Also described are plasmid for claimed method realisation, chimeric virus preparation, obtained by method described above, method of obtaining anti-serum to natural PLRV isolates and its application. Invention can be used in field of agriculture.
EFFECT: claimed is method for obtaining preparative quantities of viral particles, imitating virions of potato leaf roll virus (PLRV).
10 cl, 12 dwg
SUBSTANCE: invention offers recombinant plasmid DNA coding a chimeric antibody against human tumour necrosis factor-alpha (TNF-alpha) based on pOptiVECTM-TOPO® plasmid. Invention refers to eukaryotic cell line as a producer of antibody to TNF-alpha, method of cell line obtainment by transfection of plasmid DNA according to the invention, and method of chimeric antibody obtainment for TNF-alpha by cultivation of cell line according to the invention.
EFFECT: increased synthesis level for antibodies against TNF-alpha by producer cells.
12 cl, 8 dwg, 1 tbl, 3 ex
SUBSTANCE: invention relates to biotechnology, i.e. use of asparaginase and a method of foodstuff preparation on the basis of asparaginase. Asparaginase, which has an amino acid sequence at least 90% homologous to the amino acid sequence of <SEQ ID NO:2>, and which after an incubation period of 5 min. within the temperature range of 70°C to 100°C has a residual activity of 200 U/mg, may be used for preparing a foodstuff or a stimulant. The foodstuff preparation method involves the incubation of the foodstuff with the above mentioned asparaginase at the incubation temperature of at least 50°C. Where necessary, the foodstuffs shall be warmed up to a temperature by at least 10°C higher than the incubation temperature. Where necessary, asparaginase shall be separated from the foodstuffs or amidohydrolase shall be inactivated. If required, the separated asparaginase shall be reused.
EFFECT: invention enables the reduction of asparagine or acrylamide contents in the asparagine-containing foodstuffs.
15 cl, 5 dwg, 11 tbl, 4 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to virology and concerns the influenza virus strain A/17/New Caledonia/99/145 (H1N1) used as an attenuation donor, as well as vaccinating strains A/17/California/66/4412 (H2N2) and A/17/Tokyo/67/912 (H2N2). The presented vaccinating strains are reassortants and produced by crossing the epidemic viruses A/California/1/66 (H2N2) and A/Tokyo/3/67(H2N2) respectively with the strain A/17/New Caledonia/99/145 (H1N1). The presented strains A/17/California/66/4412 (H2N2) and A/17/Tokyo/67/912 (H2N2) are deposited in D. I. Ivanovskiy's Institute of Virology, under Nos. 2650 and 2651 respectively. The reassortants have inherited two genes coding surface virus antigen (haemaglutinin and neuraminidase) from the epidemic viruses A/California/1/66 (H2N2) and A/Tokyo/3/67(H2N2) respectively, and the rest six genes coding nonglycosylated proteins from the attenuation donor A/17/New Caledonia/99/145 (H1N1).
EFFECT: presented inventions enable producing the vaccinating strains to be used in preparing the intranasal vaccines.
3 cl, 2 tbl