Physiologically active polypeptide conjugate showing prolonged half-time index in vivo
FIELD: medicine, genetic engineering.
SUBSTANCE: invention relates to a method for preparing protein conjugates. Method involves adding immunoglobulin or physiologically active polypeptide to a single end of nonpeptide polymer having two reactive terminal groups in the presence of reducing agent. From a prepared reaction mixture the complex containing nonpeptide polymer bound with immunoglobulin or physiologically active polypeptide is isolated. Then free reactive terminal group of nonpeptide polymer of the complex is added covalently to immunoglobulin or physiologically active polypeptide in the presence of reducing agent to obtain a protein conjugate. Protein conjugate containing physiologically active polypeptide bound, nonpeptide polymer and immunoglobulin that are added one to another covalently in indicated order is isolated. Invention provides preparing a protein conjugate useful for manufacturing a polypeptide drug owing to its enhanced stability in vivo, prolonged blood half-time index and decreased immunogenic properties.
EFFECT: valuable medicinal properties of conjugate, improved preparing method.
21 cl, 16 dwg, 14 tbl, 12 ex
The text descriptions are given in facsimile form.
1. Protein conjugate containing
i) a physiologically active polypeptide;
ii) ones polymer selected from the group consisting of poly(ethylene glycol), poly(propylene glycol), copolymers of ethylene glycol-propylene glycol, polyoxyethylene polyol, polyvinyl alcohol, polysaccharide, dextran, polyvinylalcohol ether, poly(lactic-glycolic) acid, a biodegradable polymer, lipid is alimera, chitin and hyaluronic acid; and
iii) an immunoglobulin,
which are covalently linked to each other in the specified order,
and with a prolonged half-life in vivo physiologically active polypeptide.
2. The protein conjugate according to claim 1, ones where the polymer contains at both ends of the two reactive groups through which the polymer is covalently bonded to a physiologically active polypeptide and the immunoglobulin.
3. The protein conjugate according to claim 2, where the immunoglobulin is covalently bonded to at least two physiologically active polypeptide with polymer ones.
4. The protein conjugate according to claim 1, where the immunoglobulin is selected from the group consisting of IgG, IgA, IgD, IgE, IgM, and mixtures thereof.
5. The protein conjugate according to claim 4, in which the immunoglobulin is selected from the group consisting of IgG1, IgG2, IgG3, IgG4, and mixtures thereof.
6. The protein conjugate according to claim 4, where the immunoglobulin is a human immunoglobulin.
7. The protein conjugate according to claim 1, where the immunoglobulin is selected from the group consisting of immunoglobulin with glycosylation of wild-type immunoglobulin with high or low degree of glycosylation applicationvantage immunoglobulin and their combinations.
8. The protein conjugate according to claim 7, where the increase or decrease in the degree of glycosylation or glycosylamine immunoglobulin the and carry out the method, selected from the group consisting of a chemical method, enzymatic method, biotechnological methods or their combinations.
9. The protein conjugate according to claim 2, where the reactive group ones polymer selected from the group consisting of aldehyde, propionic aldehyde, butyric aldehyde, maleimide and derived succinimide.
10. The protein conjugate according to claim 9, where the derived succinimide is succinimidylester, succinimidylester, hydroxysuccinimidyl or succinimidylester.
11. The protein conjugate according to claim 9, ones where the polymer contains at both ends of the aldehyde group.
12. The protein conjugate according to claim 1, ones where the polymer is covalently bonded at the ends with end amino group, a lysine residue, a histidine residue or a cysteine residue of the immunoglobulin and end amino group, a lysine residue, a histidine residue or a cysteine residue physiologically active polypeptide, respectively.
13. The protein conjugate according to claim 1, ones where the polymer is a polyethylene glycol.
14. The protein conjugate according to claim 1, where the physiologically active polypeptide is selected from the group consisting of a hormone, cytokine, enzyme, antibody, growth factor, factor regulation of transcription factor blood, vaccines, structural proteins, ligand proteins and receptor.
15. alcovy conjugate on 14 where the physiologically active polypeptide is selected from the group consisting of human growth hormone, a hormone that stimulates growth hormone, a peptide that stimulates growth hormone, interferons, colony stimulating factor, interleukins, glucocerebrosidase, factor activation of macrophages, macrophage peptide, b-cell factor, T cell factor, protein a, allergic suppressor factor, a glycoprotein necrosis of cells, immunotoxin, lymphotoxin, tumor necrosis factor, factor, inhibiting tumor growth, transforming growth factor, α-1-antitrypsin, albumin, apolipoprotein-E, erythropoietin, erythropoietin hyperglycosylated, factor VII, factor VIII, factor IX, plasminogen activator, urokinase, streptokinase, a protein, C-reactive protein, renin inhibitor, collagenase inhibitor, superoxide dismutase, platelet-derived growth factor, epidermal growth factor, osteogenic growth factor, a protein that stimulates osteogenesis, calcitonin, insulin, atriopeptin, factor, inducing the growth of cartilage, protein activator of growth of connective tissue, follicle-stimulating hormone, luteinizing hormone, FSH-releasing hormone, nerve growth factor, parathyroid hormone, relaxin, secretin, somatomedin, insulin-like growth factor, adrenocortical oplogo hormone, glucagon, cholecystokinin, pancreatic polypeptide, gastrin-releasing peptide, corticotropin-releasing factor, thyroid hormone receptor, receptor antagonist, surface antigen cells, monoclonal antibodies, polyclonal antibodies, fragments of antibodies, including Fab, Fab', F(ab')2, Fd, scFv, and derived from a virus antigen used for production of vaccines.
16. The protein conjugate according to clause 15, where the physiologically active polypeptide is a growth factor human, α-interferon, β-interferon, granulocyte colony-stimulating factor or erythropoietin.
17. A method of obtaining a protein conjugate according to claim 1, including
A. covalent joining one end of the polymer ones with two reactive terminal groups or immunoglobulin or physiologically active polypeptide in the presence of a reducing agent;
b. selection from the resulting reaction mixture complex ones containing polymer associated with immunoglobulin or physiologically active polypeptide;
C. covalent joining of the free reactive end group ones polymer complex of immunoglobulin or physiologically active polypeptide in the presence of a reducing agent with the floor is rising protein conjugate; and
d. the selection of the protein conjugate containing a physiologically active polypeptide, polymer ones and immunoglobulin, which is covalently attached to one another in the specified order.
18. The method according to 17, where the molar ratio of the physiologically active polypeptide and the ones of the polymer phase (a) is selected from the range from 1:2.5 to 1:5.
19. The method according to 17, where the molar ratio of immunoglobulin and ones of the polymer phase (a) is selected from the range from 1:5 to 1:10.
20. The method according to 17, where the molar ratio of the complex obtained in stage (b), and physiologically active polypeptide or immunoglobulin at the stage (s) selected from the range from 1:1 to 1:3.
21. The method according to 17, where the reducing agent is cyanoborohydride sodium, sodium borohydride, dimethylaminoborane or pyridinoyl.
FIELD: biotechnology, food processing industry, derivatives of amino acids.
SUBSTANCE: invention proposes a method for enzymatic preparing α-L-aspartyl-L-phenylalanine-β-ester (α-L-(β-o-substituted aspartyl)-L-phenylalanine) from L-aspartic acid α,β-diester and L-phenylalanine at temperature 5-400C and pH 7-10 for period sufficient for formation of α-L-aspartyl-L-phenylalanine-β-ester from L-aspartic acid α,β-diester and L-phenylalanine. Proposed method is used in a method for preparing α-L-aspartyl-L-phenylalanine-α-methyl ester that comprises a step for synthesis of α-L-aspartyl-L-phenylalanine-β-methyl ester and step for conversion of indicates ester to α-L-aspartyl-L-phenylalanine-α-methyl ester. Using this invention provides synthesis of α-L-aspartyl-L-phenylalanine-β-ester followed by preparing α-L-aspartyl-L-phenylalanine-α-methyl ester by simplified and inexpensive scheme and with enhanced yield of the end product.
EFFECT: improved method of synthesis.
5 cl, 1 dwg, 4 tbl, 12 ex
FIELD: pharmaceutical chemistry and biotechnology.
SUBSTANCE: invention relates to highly purified lipopeptide antibiotic and pharmaceutical compositions comprising this compound. Invention also discloses daptomycin purification method including a series of consecutive stages of anion-exchange chromatography, hydrophobic chromatography, and anion-exchange chromatography as well as a method for purification of daptomycin using anion-exchange chromatography enhanced by modified buffer. Invention discloses improved method for production of daptomycin via fermentation of Streptomyces roseosporus followed by purification stage. Invention further discloses using high-efficiency liquid chromatography methods for checking purity of daptomycin. Disclosed are likewise lipopeptide micelles and methods for making these micelles as well as use of lipopeptide micelles in purification of lipopeptide antibiotics, such as daptomycin, and therapeutical application thereof. Invention allows preparation of lipopeptide antibiotics with 95 to 98% purity.
EFFECT: increased antibacterial activity of lipopeptide antibiotics against gram-positive microflora.
68 cl, 19 dwg, 3 tbl, 18 ex
FIELD: immunology, medicine.
SUBSTANCE: claimed recombinant antibody (Ab) has at least constant regions in heavy and light chains representing human Ab regions. Said At inhibits bonding of integrine recognizing RGD and SVVYGLR sequences to integrine or fragment thereof. Also disclosed are nucleotide sequences (NS) encoding heavy and light chains of recombinant Ab as well as expression vectors containing respective NS. Described are host cell for Ab production, transformed with two vectors for expression of Ab heavy and light chains and method for abovementioned host cell application to produce recombinant Ab. Ab of present invention is useful in diagnosis and treatment of autoimmune diseases, rheumatism and rheumatoid arthritis.
EFFECT: therapeutic methods of increased efficiency.
45 cl, 14 tbl, 28 ex
SUBSTANCE: disclosed is expression vector containing ColE1 replication system wherein homology of RNAI and RNAII of ColE1 replication initiating site with free tRNA is modified by one or more mutation in encoding region of RNAI or RNAII gene. Said mutations lead to substitution of one or more bases in loop 1, 2, 3 of RNAI or RNAII. Also described are bacterial cell transformed by said vector and method for production of desired protein from abovementioned bacterial cell. Claimed method includes cell culturing, protein isolation and purification thereof. Increased homology ratio of RNAI and RNAII with free tRNA causes increased amount of plasmida copies. Decreased homology ratio of RNAI and RNAII with free tRNA causes reduced metabolite load in cell that is highly for recombinant protein production.
EFFECT: improved method for controlling of plasmida copy amount.
12 cl, 1 dwg, 7 tbl, 2 ex
FIELD: biotechnology, in particular production of glycopeptide-originated biologically active substances.
SUBSTANCE: claimed method includes cultivation of lactic acid bacteria on nutrient medium containing defatted desiccated milk and yeast extract, followed by treatment of cultural liquid with acidic proteolytic enzymes, hydrolysis, separation of cultural liquid by centrifugation on biomass and milky whey which are treated separately. Milky whey is separated by ultrafiltration on high- and low molecular fractions. High molecular fraction of milky whey is lyophilized. Low molecular fraction of milky whey is vaporized to produce lactic also is disclosed or salts thereof. Biomass is suspended in ammonia water and separated by centrifugation on liquid and solid phases. Liquid phase is concentrated by ultrafiltration. Zinc acetate solution is added to obtained concentrated to produce zinc-containing complex. Solid phase (biomass) is treated with ultrasound or by "freezing/defrosting" method for cell wall decomposition followed by boiling, cooling, lysocim addition, and hydrolysis. Hydrolyzate is acidified with acetic acid and separated on liquid and solid phases. Solid phase obtained from hydrolyzate is dried to produce preparation containing cell wall residues and non-dissolved glycopeptide. Liquid phase obtained from hydrolyzate is fractioned by ultrafiltration on high- and low molecular fractions. High molecular fraction is lyophilized to produce preparation containing lysocim and high molecular glycopeptide. Low molecular fraction is treated by chromatography, filtered, boiled down, and lyophilized to produce preparation.
EFFECT: improved method for glycopeptide production.
3 cl, 1 tbl, 3 ex
FIELD: biochemistry, biotechnology, medicine, in particular bioactive peptides.
SUBSTANCE: latarcins are obtained from poison of spider Lachesana tarabaevi. Latarcins obtained by chemical synthesis exhibit biological properties being identical to natural ones. Said latarcins comprise of 24 amino acid residues, represent linear peptized and have no cysteine residues. Latarcins have antibacterial properties in relates to gram-positive and gram-negative bacteria and yeast fungi. Latarcin peptide has amino acid sequence of H2N-X1 1-Leu1-Lys3-Asp4-Lys5-X2 6-Lys7-Ser8-Met9-Gly10-Glu11-Lys12-Leu13-Lys14-Gln15-Tyr16-Ile17-Gln18-Thr19-Trp20-Lys21-Ala22-Lys23-Phe24-NH2, wherein X1 is glycine Gly1 or serine Ser1 residue; X2 is phenylalanine Phe6 or valine Val16 residue. Latarcins have high antibacterial activity, cytotoxicity in relates to certain cells and are useful in production of pharmaceutical agents.
EFFECT: latarcin peptides having increased antibacterial activity.
5 dwg, 3 tbl, 8 ex
FIELD: biochemistry, biotechnology, medicine, in particular bioactive peptides.
SUBSTANCE: latarcins 3a and 3b are obtained from poison of spider Lachesana tarabaevi. Said latarcins comprise of 24 amino acid residues, represent linear peptized and have no cysteine residues. Latarcins obtained by chemical synthesis exhibit biological properties being identical to natural ones. Latarcins have antibacterial properties in relates to gram-positive and gram-negative bacteria and yeast fungi. Latarcin peptide has amino acid sequence of H2N-Ser1-Trp2-X3-Ser4-Met5-Ala6-Lys7-Lys8-Leu9-Lys10-Glu11-Tyr12-Met13-Glu14-Lys15-Leu16-Lys17-Gln18-Arg19-Ala20-NH2, wherein X - lysine Lys3 residue (latarcin 3a, LtAMP-3a) or alanine Ala3 residue (latarcin 3b, LtAMP-3b). Latarcins have high antibacterial activity, cytotoxicity in relates to certain cells and are useful in production of pharmaceutical agents.
EFFECT: latarcin peptides having increased antibacterial activity.
6 dwg, 3 tbl, 8 ex
FIELD: gene and protein- engineering, medicine, pharmaceutical industry.
SUBSTANCE: invention relates to recombinant plasmide DNA pER-Gl is constructed, which provides synthesis of hybrid polypeptide containing human glucagone and intein in Escerichia coli cells. Producer of said hybrid polypeptide is obtained by transformation of E.coli ER2566 with pER-Gl plasmide. Method for production of human recombinant glucagone includes providing and cultivation of hybrid polypeptide producer containing human glucagone and intein followed by isolation and cleavage of said hybrid protein.
EFFECT: increased yield of human recombinant glucagone; simplified technology.
3 cl, 3 ex, 3 dwg
FIELD: biotechnology, biochemistry.
SUBSTANCE: invention represents enzyme that catalyzes reaction for formation of peptide from carboxyl component and amine component. Also, invention relates to strains of microorganism Empedobacter and genus Sphingobacterium that produce this enzyme, and to a method for preparing dipeptides from carboxyl component and amine component using this enzyme. Invention provides synthesis of peptide without carrying out the complex method of synthesis.
EFFECT: improved and easy method of synthesis, reduced cost, high yield of peptide.
11 cl, 18 tbl, 27 ex
SUBSTANCE: disclosed is new enzyme useful as catalyst of peptide synthesis reaction from carboxyl component and amino component. Said enzyme is produced by bacterial host cell cultivation transformed with DNA encoding peptide-forming enzyme, followed by accumulation of target enzyme. Obtained culture is blended with carboxyl component and amino component to produce dipeptide.
EFFECT: simplified method with increased yield.
38 cl, 4 dwg, 18 tbl, 39 ex
FIELD: molecular biology, biochemistry, microbiology.
SUBSTANCE: invention relates to using adenylcyclase toxin from microorganisms of genus Bordetella in producing protein vectors used in targeting a molecule to cell expressing CD11b. This molecule is chosen from the following group comprising peptides, glycopeptides, lipopeptides, polysaccharides, oligosaccharides, nucleic acids, lipids and chemicals. Molecule shows antigenic properties, it is targeted and translocated to cytosol in cells expressing CD11b. Also, invention relates to using immunogenic composition based on adenylcyclase from microorganism of genus Bordetella as a vaccine for preparing the immunotherapeutic composition. Invention describes a protein vector comprising adenylcyclase from microorganism of genus Bordatella for targeting to cells expressing CD11b, in particular, to cytosol of these cells directly. Invention provides agents that can target molecules in vivo specifically to definite populations of pAPC for providing possibility for stimulation of the immune system.
EFFECT: valuable biological properties of vectors.
56 cl, 60 dwg, 1 tbl, 1 ex
FIELD: oncology and biotechnology.
SUBSTANCE: invention concerns conjugates used for treatment of malignant tumor. Conjugate includes staphylococcal or streptococcal wild-type superantigen or modified superantigen and antibody constituent. Bacterial superantigen is modified to reduce serum reactivity with preserved its antigenic activity. Amino acid sequence of superantigen incorporates A-E regions determining binding to TCR and MHC molecules class II. Invention is directed to preparing antitumor drug and also to preparing pharmaceutical composition.
EFFECT: use of the conjugate according to invention activate immune system and, therefore, resistance of mammalian against malignant tumor.
67 cl, 11 dwg, 1 tbl, 11 ex
FIELD: biotechnology, enzymes.
SUBSTANCE: invention relates to modified gelonin enzyme and method for production thereof. Claimed method includes identification of antigen regions in enzyme by using antibody, removing of at least one antigen region and determination of fermentative activity of obtained modified gelonin enzyme. Also disclosed is multipeptide protein compound based on modified gelonin enzyme added to the second polypeptide such as antibody, toxin or other enzyme and having decreased antigenicity. Also disclosed is humanized recombinant toxin gelonin (HRTG) with decreased antigenicity and amino acid sequence wherein at least three amino acid residues from one or more 1,2,3, or 4 antigenic domains are substituted with other amino acid residues. HRTG, modified gelonin enzyme, and multipeptide protein compound are useful as drug for therapy of cancer.
EFFECT: modified proteins with decreased antigenicity and high cytotoxicity in relates to tumor cells, in particular myeloma cells.
27 cl, 11 dwg, 9 tbl, 6 ex
FIELD: biotechnology, immunology.
SUBSTANCE: invention reports about preparing and characterizing two forms of Nogo protein bound with the natural myelin with respect to the presence of immunogenic properties in their. These two forms correspond to natural products of alternative splicing, their fragments and derivatives, in particular, derivatives comprising deletions in amino acid residues, and chimeric protein also and comprising novel immunogenic polypeptides and their fragments. Invention can be used in medicine for diagnostic and curative aims.
EFFECT: valuable medicinal properties of protein.
18 cl, 79 dwg, 3 tbl, 8 ex
FIELD: medicine, biotechnology, pharmacy.
SUBSTANCE: invention relates to exchangers of ligand/receptor specificity delivering antibodies to receptors on pathogen. In particular, invention describes variants related to manufacturing and using exchangers of ligand/receptor specificity. Exchangers comprise at least one specificity domain containing ligand for receptor wherein ligand is not antibody or its part, and at least one antigenic domain combined with abovementioned specificity domain wherein antigenic domain comprises epitope of pathogen or toxin. Advantage of the invention involves enhanced specificity in delivery of drug.
EFFECT: improved and valuable properties of exchangers.
30 cl, 5 tbl, 6 ex
FIELD: biotechnology, medicine.
SUBSTANCE: the present innovation deals with CFP10-ESAT6 hybrid protein that is able to induce Mycobacterium tuberculosis-specific hypersensitivity reaction of delayed type. This hybrid protein contains complete protein CFP10 out of M.tuberculosis combined with complete protein ESAT6 out of M.tuberculosis through linker amino acid sequence. It is, also, [presented a recombinant plasmid DNA pTBD16 for expression of hybrid protein CFP10-ESAT6. The latter should be obtained due to applying DLT1270 E.coli strain transformed with the obtained recombinant plasmid DNA pTBD16. The innovation suggests to apply the above-mentioned protein as a diagnostic kit of tuberculous infection both in mammalians and human beings. The method for predicting tuberculous infection has been revealed. Tuberculous infection should be predicted due to induction of hypersensitivity reaction of delayed type in case of subcutaneous injection of hybrid protein. Application of the present innovation enables to obtain a Mycabacterium tuberculosis-referring diagnostic kit, at specificity being 100%, sensitivity - 80%, not less.
EFFECT: higher accuracy and efficiency of diagnostics.
6 cl, 4 dwg, 4 ex
FIELD: biotechnology, molecular biology, medicine.
SUBSTANCE: invention discloses amino acid sequences of human obesity polypeptide (OB) two isoforms possessing capacity for modulation of animal body mass, their signal peptide-containing precursors and analogues. Polypeptide isoforms are prepared as result of insertions, deletions and amino acid changes that retain activity typical for nonmodified forms of OB-polypeptides, and polyclonal and monoclonal antibodies interaction specifically with new agents modulating the body mass value also. Invention describes DNA sequences encoding these polypeptides and their analogues, vector structures comprising these sequences used for preparing recombinant forms of OB-polypeptides. Invention proposes using new polypeptides and their analogues as an active component in pharmaceutical compositions. Using this invention can promote to solving the problem for providing medicine, veterinary science and animal husbandry with effective agent used for decreasing the body mass value. Invention can be used in medicine for diagnosis and treatment of pathological states associated with disturbance of regulation of human body mass, and in animal husbandry and veterinary science.
EFFECT: valuable biological, medicinal and veterinary properties of polypeptide.
23 cl, 71 dwg, 12 tbl, 17 ex
FIELD: biotechnology, gene engineering, microbiology.
SUBSTANCE: invention relates to TUL4spCBD recombinant protein comprising TUL4 Francisella tulergenis protein, Gly-Ser spacer and cellulose-binding domain of CelD endoglucanase gen from Anaerocellum thermophilus. Said protein has TUL protein antigen properties and is capable of spontaneous binding to cellulose-containing sorbents. Described is recombinant plasmid pTUL4spCBD DNA of 4388 n.p. length, encoding TUL4spCBD recombinant protein. Abovementioned plasmid contains: artificial bacterial operon of TUL4spCBD recombinant protein, including promoter region of N5 bacteriophage earlier promoter (7-87 n.p.); TUL4spCBD recombinant protein gene (115-1113 n.p.); transcription terminator (1134-1230 n.p.); beta-lactamase bacterial operon (4183-3323 n.p. of complementary chain)), providing ampicillin resistance; ColE1-type bacterial site of replication initiation, providing plasmid replication in E.coli strain (4388 n.p.). Also discoised is M15[pREP4, pTUL4spCBD] Escherichia coli strain as producer of TUL4spCBD recombinant protein and method for production of purified, concentrated and immobilized TUL4spCBD recombinant protein from said strain by treatment of cell hydrolyzate supernatant from M15[pREP4, pTUL4spCBD] Escherichia coli with cellulose-containing sorbent. Method for production of specific antibodies against TUL4spCBD protein also is described. Said method includes animal immunization with TUL4spCBD recombinant protein immobilized onto cellulose.
EFFECT: method for high yield production of TUL4spCBD recombinant protein; simplified method for purification, concentration and immobilization of said protein.
5 cl, 1 dwg, 4 ex
FIELD: genetic engineering, pharmaceutical and medical-biological industry.
SUBSTANCE: invention proposes a chimeric sequence of nucleic acid encoding a fused polypeptide able to bind with the vessel endothelium growth factor (VEGF) and to inhibit its specific mitogenic effect. The fused polypeptide molecule comprises immunoglobulin-like domain 2 of VEGF-receptor Flt1, immunoglobulin-like domain 3 of VEGF-receptor Flk 1 or Flt4 and multimerizing component represented by either domain Fc IgG or heave chain IgG. By expression of the proposed chimeric sequence or its two successively joined copies in a host-cell a monomer or dimer of the fused polypeptide are prepared, respectively, that can be used for suppression of VEGF activity in mammals, in particular, in humans. New VEGF inhibitors differ from the known one by the improved pharmacokinetics.
EFFECT: improved preparing method, valuable biological properties of polypeptide.
23 cl, 67 dwg, 1 tbl, 35 ex
FIELD: biotechnology, microbiology, genetic engineering.
SUBSTANCE: invention proposes a new recombinant plasmid pR752 (5269 pair bases) comprising genetic construction under control of bacteriophage T5 promoter and encoding a module polypeptide consisting of 6 histidine residues, hemoglobin-like protein of E. coli, modified fragment of large T-antigen SV-40, translocation domain of diphtheria toxin, spacer sequence (Gly-Ser)5 and human epidermal growth factor (6 His-HMP-NLS-Dtox-(Gly-Ser)5-EGF) and designated for the directed transfer of photosensitizers into target-cell nuclei. By transformation of the strain E. coli M15 (rep 4) with plasmid pR752 the recombinant strain E. coli VKPM B-8356 as a producer of new polypeptide vector is prepared. The usage of this new strain is able to enhance the effectiveness of effect of photosensitizers by some orders. Invention can be used in medicinal-biological industry in preparing agents providing the directed transport of photosensitizing agents into tumor cell nuclei.
EFFECT: valuable biological and medicinal properties of polypeptide.
3 dwg, 4 ex
FIELD: immunology, biotechnology.
SUBSTANCE: invention describes murine antibody and its humanized variant (CDP870) showing specificity to human tumor necrosis factor-alpha. Amino acid sequence is given in the description. Also, invention describes compounds showing affinity with respect to human tumor necrosis factor-alpha based on humanized antibody wherein lysylmaleimide group bound covalently with one or some methoxypoly(ethylene glycol) molecules by lysyl residue is joined to one of cysteine residues by C-end of heavy chain of the humanized antibody. Invention discloses DNA sequences encoding antibodies showing specificity to human tumor necrosis factor-alpha and variants if expression vectors involving indicated DNAs. Also, invention describes variants of a method for preparing a host-cell using expression vectors and variants of a method for preparing antibodies based on prepared host-cells. Invention discloses therapeutic compositions used in treatment of pathology mediated by tumor necrosis factor-alpha based on antibodies. Invention provides providing antibodies showing high affinity: 0.85 x 10-10 M for murine antibodies and 0.5 x 10-10 M for its humanized variant and low immunogenicity for human for humanized antibodies. Part of patients with improved ACR20 in administration of 5 and 20 mg/kg of CDP870 is 75% and 75% in 8 weeks, respectively. Half-life value of CDP870 in plasma is 14 days.
EFFECT: valuable biological and medicinal properties of antibodies.
58 cl, 24 dwg, 6 tbl, 1 ex