Photosensitive chimeric protein gpcr

FIELD: biotechnology.

SUBSTANCE: invention relates to production of a photosensitive chimeric protein capable of incorporating a light signal into the signalling cascade of metabotropic glutamate receptor 6 (mGluR6), which is a natural component of the ON-bipolar cell membrane in the inner layer of the retina, which can be used in medicine. A GPCR chimeric protein is obtained comprising domains of at least two members of a protein receptor superfamily conjugated with G proteins (GPCR), a nucleic acid encoding the said protein, an expression vector comprising the said nucleic acid, as well as a transgenic cell line containing genetic information, encoding a chimeric GPCR protein.

EFFECT: invention allows effective drug therapy and production of an effective drug for vision improvement, in particular, for treatment of vision loss due to degeneration of retinal photoreceptors.

32 cl, 6 dwg, 1 tbl, 1 ex

 



 

Same patents:

FIELD: medicine.

SUBSTANCE: claimed invention relates to biotechnology and represents an expression plasmid without resistance to an antibiotic, containing a polynucleotide, coding a repressor protein cI. The expression of the said repressor protein regulates the expression of a toxic gene product, embedded into a non-essential section of a host genome. The claimed invention also discloses a constructed host cell, belonging to Gram-negative bacteria, which contains the said plasmid. The host cell is used to obtain the plasmid or to obtain a protein or an immunogen, if the expression plasmid additionally contains genes, coding the said protein or immunogen. The method of obtaining the expression plasmid and the method of obtaining the protein or immunogen are carried out in several stages. First, a strain of host cells, belonging to the Gram-negative bacteria, is created by embedding by the allele substitution of the gene, coding the toxic product, into the non-essential section of a host chromosome. After that, construction of the DNA-plasmid, which contains the gene, coding the repressor protein cI, and in case of necessity the gene, coding the protein or immunogen, is performed. Then, the obtained host cells are transformed by the said plasmid and grown in the presence of sucrose at a temperature of 30-42°C.

EFFECT: invention makes it possible to realise control of the toxic gene, localised on a chromosome, by means of the repressor, localised on the plasmid in the absence of selective pressure by an antibiotic.

33 cl, 31 dwg, 8 tbl, 10 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to biochemistry and represents a new bifunctional PSH protein containing human peroxiredoxin Prx6 and manganese superoxide dismutase MnSOD possessing the antioxidant activity of superoxide dismutase and peroxidase. What is also described is a chimeric nucleic acid coding the presented protein. A method for preparing the presented protein by culturing cells of the strain E.coli BL21(PSH) transformed by constructed recombinant expression vector based on pET22b(+) plasmid is disclosed.

EFFECT: invention enables producing high-yield protein PSH possessing the high antioxidant activity.

4 cl, 6 dwg, 3 tbl, 7 ex

FIELD: chemistry.

SUBSTANCE: invention relates to the field of biotechnology. Claimed is a recombinant plasmid pCHBH for an expression of a sequence, coding procarboxypeptidase B in Pichia pastoris, which has a size of10466 t.b.p. and consists of XhoI/EcoRI -fragment of DNA of vector pP1C9K with a size of 9246 t.b.p. and XhoI/EcoRI -fragment of DNA with a size of 1220 t.b.p., including a region, coding the signal peptide of α-factor of Saccharomyces cerevisiae,anda synthetic gene of human procarboxypeptidase B with a nucleotide sequence, corresponding to the sequence, represented on fig. 2. In addition, a recombinant strain of Pichia pastoris GS115CPBH - producent of human procarboxypeptidase, obtained as a result of the parent strain transformation by the said plasmid, is described.

EFFECT: invention makes it possible to obtain human procarboxypeptidase and a corresponding to it active form in an increased quantity, constituting not less than 9,0 units/ml in comparison with the prototype.

2 cl, 4 dwg, 4 ex

FIELD: chemistry.

SUBSTANCE: invention relates to the field of biochemistry, in particular to recombinant factor VIII, which contains one or more mutations, resulting in an increased stability of both the factor VIII and factor VIIIa, as well as to a pharmaceutical composition for treating haemophilia containing it. Also described is a molecule of nucleic acid, coding the said recombinant factor VIII, and an expression vector and host-cells, containing the said molecule of nucleic acid. The invention also relates to a method of obtaining the said factor VIII, as well as to its application in the method of treating haemophilia A in an animal.

EFFECT: invention makes it possible to obtain a biologically active factor VIII with an increased stability.

50 cl, 12 dwg, 5 tbl, 9 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to biochemistry, in particular to a monoclonal human antibody, specific to alpha-toxin of S. aureus. The claimed invention additionally relates to pharmaceutical compositions for treatment of prevention of the abscess formation in an organ, which contains at least one antibody or one nucleic acid, which codes the said antibody.

EFFECT: invention makes it possible to extend an assortment of antibodies, specific to alpha-toxin of S aureus.

23 cl, 7 dwg, 4 tbl, 6 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to biotechnology, namely to leukolectins, and can be used in medicine. What is prepared is the polypeptide leukolectin characterised by SEQ ID NO:1-8. The recombinant preparation is ensured by using a nucleic acid coding it and integrated into an expression vector which is used to transform a host cell. Testing absence-presence or determining an amount of the polypeptide leukolectin are ensured by using an antibody or an antigen-binding fragment of a variable region of the above antibody which is specifically bound to the polypeptide leukolectin. The polypeptide leukolectin or the nucleic acid coding it are used as ingredients of a pharmaceutical composition in therapy of pathological disorders of skin and mucous membranes.

EFFECT: invention enables treating or preventing autoimmune disorders of skin, inflammatory diseases of skin or mucous membrane, or injured skin in an animal effectively.

16 cl, 19 dwg, 3 tbl, 12 ex

FIELD: biotechnologies.

SUBSTANCE: group of inventions refers to biotechnology and deals with new nucleotide sequences of Torque teno virus (TTV) and vectors containing such sequences. Extracted polynucleotide molecule contains polynucleotide sequence chosen from the group consisting of SEQ ID NO:4, sequence complementary to SEQ ID NO:4 and polynucleotide that is at least 95% identical to SEQ ID NO:4.

EFFECT: inventions can be use for production of vaccines to prevent diseases of pigs and other animals, which are caused by Torque teno virus.

4 cl, 7 dwg, 3 tbl, 10 ex

FIELD: biotechnologies.

SUBSTANCE: recombinant nucleic acid expresses one or several polypeptides of interest, a vector of expression and bacteria, which contain this recombinant nucleic acid. The recombinant nucleic acid contains a natural promotor of a gene of HU-like DNA-binding protein (PhilA) of Lactococcus type with the sequence SEQ TD NO:28, or its homological or functional version, which at least by 95% identical to the promotor with sequence SEQ ID NO:28, functionally linked with one or several open reading frames, heterological for the promotor RhIIA, where the promotor RhIIA is located above one or several open reading frames. The expression vector contains the above recombinant nucleic acid, preferably, the specified vector is produced from pTINX. A bacterium contains the above recombinant nucleic acid or the above vector.

EFFECT: proposed invention makes it possible to increase level of expression of polypeptide genes of interest and therefore produce sufficient number of expressed proteins.

19 cl, 26 dwg, 12 tbl, 9 ex

FIELD: biotechnologies.

SUBSTANCE: proposed vector is designed on the basis of a vector plasmid pEGFP-Nl, containing a DNA fragment, which codes a promotor of a heat shock protein gene hsp70 Drosophila melanogaster and a regular sequence upstream, containing heat shock elements (HSE) in different quantities, a polylinker zone, a gene of green fluorescent protein (GFP) and a gene of stability to neomycin, at the same time the promotor is capable of activation under action of temperature of heat shock of mammals or under toxic effect. Such vector is activated as temperature increases to 38°C or in case of toxic impact at transgenic cells or tissue of mammals. Activity of a promotor within the vector may be regulated, i.e. it is possible to cause either its hyperactivity or its weak leak, or to block activity of this promotor by saturation of the regulatory area with HSE elements.

EFFECT: invention may be used to produce transgenic preparations, where an investigated or used gene will be under control of a regulated non-viral promotor.

3 cl, 20 dwg, 2 ex

FIELD: biotechnologies.

SUBSTANCE: invention relates to a molecule of nucleic acid, which is a cyclic or a linear vector fit for expression, of at least one target polypeptide in cells of mammals, including (a) at least one expressing cassette (POI) for expression of the target polypeptide; (b) an expressing cassette (MSM), including a gene of a selective marker of mammals; (c) an expressing cassette (MASM), including an amplificated gene of a selective marker of mammals; besides, the expressing cassette (POI) is flanked in direction 5' by the expression cassette (MASM), the expression cassette (MSM) is localised in direction 3' from the expression cassette (POI) and in which the expression cassettes (MASM), (POI) and (MSM) are arranged in the same orientation from 5' to 3'. Also the method is disclosed to produce the specified molecule of nucleic acid of the vector, as well as a cell of a host mammal, containing the specified molecule of nucleic acid of the vector, the method to produce a host cell containing the specified molecule of nucleic acid of the vector, and also the method to produce the target polypeptide, using the specified host cell.

EFFECT: invention makes it possible to efficiently produce a target polypeptide in mammal cells.

24 cl, 2 dwg, 4 tbl, 13 ex

FIELD: chemistry.

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.

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

FIELD: chemistry.

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

FIELD: biotechnology.

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

FIELD: medicine.

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

Siglec-15 antibody // 2539790

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

FIELD: chemistry.

SUBSTANCE: invention relates to conjugates of natriuretic compounds. In particular, conjugated forms of hBNP are proposed, which contain at least one modifying group attached thereto. The modified natriuretic compound conjugates retain activity for bonding with NPR-A receptor for stimulating cGMP production, and have longer half-life in blood flow compared to unmodified counterpart natriuretic compounds.

Class 1: Non-hydrolysable - conjugate medicinal agent remains unchanged Alky inside or PAG inside

Class 2: MicroPAGylated - the alkyl part remains splits in vivo

Class 3: Fully hydrolysable - the entire oligomer splits in vivo

= amphiphilic oligomer Nobex,

=bBNP.

EFFECT: pharmaceutical compositions which contain the disclosed conjugates and use of the disclosed conjugates to make medicinal agents.

13 cl, 13 dwg, 3 tbl, 30 ex

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