Plasminogen and plasmin variants
SUBSTANCE: invention relates to biotechnology and can be used for production of plasminogen and plasmin variants. Version of plasminogen is obtained, containing activation site and catalytic domain, in which catalytic domain contains valine, substituted for isoleucine, in position 1 of catalytic domain of human plasmin or in position, corresponding to such in catalytic domain of plasmin, which differs from human plasmin where said catalytic domain of human plasmin starts with amino acid valine in position 1, which is the same amino acid valine in position 562 Glu-plasminogen of human with SEQ ID NO: 1.
EFFECT: invention enables to obtain autoproteolytically stable variant of plasmin.
19 cl, 14 dwg, 3 tbl, 2 ex
SUBSTANCE: invention is immobilised on a matrix streptokinase which activates plasminogen to plasmin. Such streptokinase is resistant to splitting by plasmin in comparison with the corresponding streptokinase of wild type and comprises an amino acid sequence having asparagine in positions corresponding to positions 85 and 412 in SEQ ID NO:1. The invention also relates to the matrix immobilised by the said streptokinase, the method of production of plasmin, and the set for production of plasmin.
EFFECT: invention enables to maintain the functional characteristics of streptokinase while simultaneous increasing resistance to splitting by plasmin while its immobilisation on the matrix.
8 cl, 7 dwg, 8 ex
SUBSTANCE: invention relates to biotechnology and can be used to extract plasminogen or plasmin. A mixture containing fibrinogen and plasminogen or plasmin is deposited onto a chromatographic column containing an insoluble affinity chromatography matrix which is covalently cross-linked with tranexamic acid. Plasminogen or plasmin is then eluted from the affinity chromatography matrix with an aqueous solution containing a sufficient amount of a ligand which competes with plasmin or plasminogen for binding sites of said matrix which is bonded with tranexamic acid. Tranexamic acid is covalently bonded with said matrix through a linker which lies between the matrix and tranexamic acid and whose length is more than three carbon atoms.
EFFECT: invention increases efficiency of extracting plasminogen or plasmin from a mixture in the presence of fibrinogen.
10 cl, 1 dwg, 13 tbl, 6 ex
SUBSTANCE: method for assaying microorganisms for fibrinolytic activity in patients with urogenital and suppurative septic diseases, consists in measuring pathogenicity factors of microorganisms in vitro with using a preparation of lyophilised citrated rabbit plasma as a main component.
EFFECT: method is characterised by consumable availability that enables the studies researches in the clinical microbiology laboratories of healthcare facilities.
SUBSTANCE: genetically engineered technique is applied to produce in Escherichia coli cells the recombinant polypeptides with human plasminogen properties, containing a catalytic plasminogen domain, 5th plasminogen kringle domains, and a modified fragment of an amino acid sequence preceding to the 5th plasminogen kringle domain.
EFFECT: invention allows producing the polypeptides of high fibrinolytic human plaminogen activity and high yield in expression in Escherichia coli cells.
7 cl, 1 dwg, 11 ex
SUBSTANCE: genetically engineered technique is applied to produce in Escherichia coli cells the recombinant polypeptides with human plasminogen properties containing a catalytic plasminogen domain, 5th and 4th plasminogen kringle domains, and a modified fragment of an amino acid sequence preceding to 4 plasminogen kringle domains.
EFFECT: invention allows producing the polypeptides of high fibrinolytic human plaminogen activity and yield 40-60 mg per 1 l of the culture in expression in Escherichia coli cells.
7 cl, 1 dwg, 3 ex
SUBSTANCE: invention relates to production of highly-purified highly-active plasminogen-activating enzyme preparations, which may be used in biochemistry and medical science. Method implies the following: Morikrasa enzyme preparation is dissolved in tris-chloride buffer with pH 7.2-7.5 to obtain concentration of at least 2.5%. Resulting solution is fractionated by means of ion-exchange chromatography on DEAE-Toyoperl sorbent. Proteinase is eluted with NaCl concentration gradient of 0.1-1.5 M. Then, resulting solution of plasminogen activator is concentrated and demineralised by ultrafiltration using a membrane permeable for peptides with molecular weight up to 10 kDa. Plasminogen activator is purified by gel chromatography on Sephadex G 50 in 0.1 M ammonium-acetate buffer with pH 6.3-6.5, which contains 1.0-1.2 M of sodium chloride.
EFFECT: simplified method of production and shorter process time.
3 dwg, 1 tbl
FIELD: chemistry, biochemistry.
SUBSTANCE: invention refers to biochemistry and can be used in pharmaceutical industry for mixture purification of natural mixtures containing plasminogen and fibrinogen from plasminogen. Plasminogen is removed from mixture containing plasminogen and fibrinogen using insoluble chromatography matrix that is covalently cross-linked with tranexamic acid through amino group with linker of length exceeding three carbon atoms. Therefore mixture mentioned above is applied on chromatographic column containing specified insoluble matrix. Then column is washed with neutral solution containing salts, while unbound material is collected.
EFFECT: extended technical feasibilities of purification of natural mixtures containing plasminogen and fibrinogen from plasminogen thus keeping original amount of fibrinogen in mixture.
15 cl, 1 dwg, 13 tbl, 6 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
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: 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, 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: 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
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology and immunology. Described are various variants of anti-IL-1R1 antibodies. The disclosed antibodies can be applicable for treating IL-1R1-mediated disorders, including rheumatoid arthritis, asthma, and chronic obstructive pulmonary disease (COPD).
EFFECT: presented group of inventions can be used in medicine.
21 cl, 14 dwg, 12 tbl, 5 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
FIELD: medicine, pharmaceutics.
SUBSTANCE: presented invention refers to immunology. There are presented versions of antibodies neutralising a subtype group 1 and subtype group 2 influenza A virus infection. The antibody is characterised by: either a set of 3 CDR of a light and 3 CDR of a heavy chain, or the presence of variable regions of the light and heavy chains. There are disclosed: a nucleic acid molecule coding the antibody; a cell expressing the antibody; as well as a method for the attenuation of the influenza A virus infection or reducing a risk thereof with the use of the antibody in a therapeutically or preventatively effective amount.
EFFECT: using the invention provides the antibodies neutralising the influenza A virus, which can find application in medicine in treating subtypes H1, H2, H3, H5, H7, H9 influenza.
18 cl, 6 tbl, 2 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: inventions deal with infectious molecule of nucleic acid, coding infectious porcine Torque teNO viruses (PTTV), which contains at least one copy of genome sequence, selected from the group, consisting of sequences, corresponding to genotypes or subtypesPTTV1a-VA, PTTV1b-VA, PTTV2b-VA and PTTV2c-VA, as well as to biologically functional plasmid or viral vector, containing such infectious nucleic genome sequence, and host-cell, containing such plasmid or vector. In addition claimed inventions include live, attenuated expressible with vector application and purified recombinant capsid subunit or killed viral vaccines for protection against PTTV infection, as well as methods of immunisation of pigs against PTTV viral infection by said vaccine introduction.
EFFECT: characterised inventions can be used to prevent infection, caused by porcine Torque teNo virus.
23 cl, 53 dwg, 5 tbl, 24 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: method includes treating a sample with a denaturing buffer solution, sorbing short RNA on a fibre glass sorbent in the presence of a chaotropic agent, followed by washing off non-bonded biopolymers and chemical reagents from the sorbent and eluting the end product. The biological fluid sample undergoes two-step denaturing, wherein at the first step two volumes of the denaturing buffer solution are added to the sample and at the second step two volumes of 96% ethanol and equal volume of chloroform are added to the obtained mixture, followed by stirring and separating the residue by centrifuging. Non-bonded biopolymers are washed off from the sorbent twice with the buffer solution and elution of the end product from the sorbent is carried out with the buffer solution.
EFFECT: simple method, short duration of the method and high output of short RNA.
3 cl, 3 tbl, 4 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: present invention refers to biotechnology and represents a conjugate used for siRNA intracellular delivery and containing siRNA, which is conjugated by a covalent bond with a hydrophilic compound on one side, e.g. PEG, and with a hydrophobic compound, e.g. cholesterol, on the other side. By self-assembly, the conjugates are able to form homogenous nanoparticles, micellas, wherein the hydrophobic compounds are packed inside the micella; siRNA - between the hydrophobic and hydrophilic compounds, and the hydrophilic compounds - outside. The present invention also discloses methods for producing the above conjugate, pharmaceutical compositions containing the above nanoparticles for the gene therapy of various diseases depending on specific siRNA delivered. What is also disclosed is a pharmaceutical composition for treating cancer, which contains the nanoparticles containing survivin-specific siRNA.
EFFECT: invention enables increasing the siRNA stability in a living body, providing thereby the effective delivery of therapeutic siRNA into cells and the shown activity of siRNA even in a low dose of a relatively low concentration.
25 cl, 19 dwg, 1 tbl, 6 ex
SUBSTANCE: invention relates to biochemistry, particularly to methods of producing a plant with higher drought and salt tolerance compared to the wild-type plant by reducing expression/function of the protein transcription factor in the plant. The invention also relates to a plant with high drought and salt tolerance obtained using said method.
EFFECT: invention enables to efficiently obtain a plant with high drought and salt tolerance compared to the wild-type plant.
6 cl, 6 dwg, 1 tbl, 6 ex