Method of obtaining collagen from biological material
SUBSTANCE: invention represents a method of obtaining collagen from biological material, which includes milling of a raw material, liquid processing of the biological material with obtaining a collagen-containing substance, separated into a sediment and liquid fraction, characterised by the fact that as the biological material applied is a medusa, preferably Rhopilema, preferably its cupola, which is crushed preferably to 1-2 mm, and for obtaining the collagen-containing substance the material prepared in such a way is mixed with drinking water with a ratio by the raw material weight to water as 1:2 and extracted at a temperature preferably of 15-18°C for 6-12 hours with periodical mixing, after that, the obtained extract is separated into the liquid fraction and collagen-containing sediment, which is after that dehydrated to moisture weight in it not more than 10%, after which it is pre-packed and packed.
EFFECT: extension of an arsenal of methods for obtaining neutral collagen.
The invention relates to methods of producing collagen from biological material.
A method of obtaining collagen by grinding the biological material, processing naydennym solution of sodium sulfate containing 10% sodium hydroxide solution, with exposure times from 3 to 5 days, neutralizing the alkali aqueous solution of boric acid, washing with running water, dissolution of the treated material in odnomestnoi acetic acid and purification by dialysis against distilled water for 3 days (see Halkin A. M., Schechter A. B., Istrana L. P., Lemeni C. L. Collagen and its application in medicine, 1976. - M.: Medicine. - S. 47-50).
The disadvantage of this method is the long time required for separation of the finished product. In addition, to obtain collagen requires expensive reagents.
Also known is a method of obtaining collagen from biological material, comprising the grinding of raw materials, liquid handling biological material with obtaining collagen containing substance that is separated into sludge and the liquid fraction (see EN No. 2322249, 2008).
However, the use as solvent of acetic acid, causing acid reaction end product, and the presence of excessive content of acetic acid and its salts, high (20%) concentration of caustic is atra, long-term (within 24 hours) washing the treated material have the effect of restricting the application of the method. In addition, these methods are based on a relatively limited range of input sources.
The task of the invention is to provide opportunities for neutral collagen and the expansion of raw materials for its production.
The technical result - the possibility of obtaining neutral collagen and the expansion of raw materials for its production.
The problem is solved due to the fact that the method of producing collagen from biological material, comprising the grinding of raw materials, liquid handling biological material with obtaining collagen containing substance that is separated into sludge and a liquid fraction, characterized in that as the use of biological material jellyfish, preferably Rhopilema, preferably its dome, which is preferably crushed to 1-2 mm, to obtain collagen containing substance thus prepared material is mixed with potable water at a ratio by mass of the raw material to water as 1:2 and extracted within 6-12 hours at room temperature, after which the extract obtained separated into a liquid fraction and the precipitate, which is then dehydrated to the mass of moisture in it no more than 10%, after CEG is Packed and Packed. In addition, the liquid fraction of the extract used for the production of drinks.
Rhopilema asamushi is considered the most valuable of Medusa fished for food purposes 12 species scyphoid gellately. Currently, this jellyfish is implemented in Europe and South-East Asia, mainly in the form of salted-dried semi-finished processed various alum used as preservatives to increase the shelf life of the product and stabilizers of its structure. In Russia jellyfish as food is not widely used. Foreign methods of preparation and use of jellyfish in food are not acceptable for the Russian consumer due to the use of prohibited sanitary-hygienic standards for food additives in the manufacture of prefabricated jellyfish.
The basic way to handle jellyfish squad Rhizostomeae abroad [Goy J., Toulemont A. Meduses. - Monako: Musee oceanographique, 1997. - 160 p.] is the production of salted-dried products. Abroad dried and salted-dried products from jellyfish is in high demand and has a high retail price. Product eaten salted-dried as snacks with soy sauce, fried, added to salads and so on
Food facilities scyphoid jellyfish as important types of non-raw materials are the most valuable food products due to the prohibited content in them is well digestible and full of protein, biologically valuable fat, a variety of vitamins, minerals and biologically active components.
This is confirmed by histological and microstructural studies. Fabric jellyfish contains a large number of biologically valuable nucleic acids that increase the immunity of the human body. Protein jellyfish presents biologically and pharmaceutically valuable collagen - gelatinous protein, the main organic constituent of connective tissue and the organic component of bone, as evidenced by the effectiveness in the treatment of arthritis [Hsieh, Y-H. P., Leong F.-M. & J. Rudloe Jellyfish as food // Hydrobiology. 2001. V. 451. P. 11-17]. Between collagen and elastic fibers in the matrix of the mesoglea extracellular matrix are biologically active fibroblasts - the cells that produce collagen fibers in the places of their injuries.
Protein jellyfish well balanced amino acid composition, containing all the vital essential amino acids, as well as a wide set of free amino acids, which have beneficial effects on the metabolic processes in the body that characterizes the biological and nutritional value of products from the Pacific gellately.
Research fractional and fatty acid composition of lipids jellyfish suggests that lipids umbrella part and the mouth of the blades jellyfish which contain triglycerides, phospholipids, di - and monoglycerides, sterols, esters of sterols and free fatty acids.
In the fatty acid composition of the Pacific scyphoid jellyfish are saturated, monounsaturated and polyunsaturated fatty acids. The study of fatty acids found in covalently bound form in the lipid composition of different classes, suggests that lipids Pacific jellyfish have a higher biological value, as well as fatty acids 16-20% consist of a series of essential, necessary for normal functioning of the body polyunsaturated acids: linoleic, linolenic, arachidonic, eicosapentaenoic.
The mineral composition of jellyfish presents a wide range of macro - and micronutrients, predominant among which are iron, chlorine, iodine, sodium, chromium, aluminum, fluoride, manganese, and bromine.
The cost of jellyfish in Russia, below the cost of sea fishery (fish and valuable seafood).
Comparative analysis of the characteristics of the claimed solution with the characteristics of the prototype and analogues demonstrates compliance of the claimed solution to the criterion “novelty”.
The characteristics of the characterizing portion of the claims, solves the following functional tasks
Signs "as the use of biological material jellyfish" allow to draw for the production of new collagen is type of biological material, the resource which is virtually unlimited.
Signs indicating the use of Medusa preferably Rhopilema", enable the use of species of jellyfish, a significant period of use in food which is in the Eastern cooking confirmed its edibility.
Signs indicating the use of a preferably dome jellyfish, provide the possibility of involvement in the trafficking of waste jellyfish, because in the East the kitchen is used preferably tentacles.
Signs indicating that the dome is ground preferably 1-2 mm, enable rapid extraction of collagen containing materials from the crushed material by increasing the specific surface area of its particles.
Signs indicating that to obtain a collagen containing substance chopped biomaterial "is mixed with potable water at a ratio by mass of the raw material to water as 1:2 and extracted within 6-12 hours at room temperature, simplify the process of biomaterial, avoid the use of reagents and provide for effective disposal of liquid wastes from this process (due to the use for the manufacture of beverages).
Signs indicating that "the extract obtained is separated into a liquid fraction and a residue will cause separation of the extract on the departure of the (liquid fraction) and the intermediate product, containing collagen.
Signs indicating that sediment "then dehydrate until the mass of moisture in it no more than 10%", ensure the implementation of the last stage of obtaining collagen.
The signs are that the finished product is Packed and Packed"to provide the "giving" product presentation.
Signs of the second claim of the invention provide for the disposal of liquid waste production of collagen to get a useful product.
For making use of collagen jellyfish raw (TU 9250-327-00472012-09) or frozen jellyfish (TU 9265-330-00472012-2010).
The raw materials used and materials on hygiene requirements food safety must comply with the Uniform sanitary and epidemiological and hygienic requirements for goods subject to sanitary and epidemiological supervision (control), and be accompanied by documents confirming its quality.
Water used for technological purposes, must comply with SanPiN 188.8.131.524-01 "Drinking water. Hygienic requirements to water quality of centralized drinking water supply systems. Quality control".
Allowed to use the marine clean water that meets the requirements of SanPiN 2.3.4.050-96.
All technological equipment used in the manufacture of collagen, must match the Tr is the requirements SanPiN 2.3.4.050-96 "enterprises of food and processing industry (technological processes, raw materials). Production and sale of fish products. Sanitary rules and norms".
Reception of raw materials, storage until processing is carried out in accordance with the requirements of normative documents.
Received for processing jellyfish raw washed of impurities clean flowing seawater or potable water to a temperature not higher than 15°C. the Washed jellyfish sent for cutting. Cutting jellyfish carried out manually, as follows: jellyfish cut off the dome and rapalee (tentacles). The dome jellyfish make an incision lengthwise and remove the innards. The large dome is cut into pieces no larger than 5×5×7 see the Dome and/or pieces of the dome and rapalee sent for washing. The dome and/or pieces of the dome, rapalee separately each type of raw material is placed in a wooden or plastic container lined with plastic wrap, and washed 2-3 times from contamination by a stream of drinking water, allowed to use clean sea water temperatures ranging from 15 to 20°C. After draining water, the product is sent for crushing.
Frozen jellyfish thawed to a temperature of from 0 to -2°C within a block of the material after washing sent for cutting, as above.
Raw materials (including thawed) are ground in an electric grinder with a hole diameter of 1-2 mm can be used homogenizers and devices thin ismel the treatment of materials (a & m) according to their instructions.
To the crushed raw materials add drinking water in the ratio 1:2, the mixture was mixed thoroughly and leave for extraction at a temperature of 15-18°C for 6-12 hours, while periodically stirring.
Upon completion of the extraction step carry out the separation of the extract, which is divided into the sediment and the liquid part using the separator of known construction, for example I-as-2ZH when the rotational speed of 6500 rpm according to his instructions.
At the completion of phase separation carry out the drying and grinding of the resulting sludge. The process is produced in a known manner, a vacuum drying apparatus of the drum type. The drying process begins with the heating of the vacuum pot within 30 minutes the Temperature is 70°C. Then loads the received material into the apparatus in the amount of 20 kg Sunroof boiler seal, include a vacuum pump (residual pressure of 2 kPa). The temperature inside the boiler is 35°C. Include rotation of the boiler, the angle of rotation should not exceed 360°. After 3.5 h of drying stops the rotation of the boiler, relieve the vacuum in the boiler load grinding media for grinding product. Drying continued for 2.5 h with the same parameters. Then the temperature was lowered to 50°C and continue the drying process even 2,0 hours Then the rotation of the boiler remains the pour, relieve the vacuum, open the hatch of the boiler, in its Central part enter the sieve and the collection of the finished product. The hatch closed, the boiler is again put into rotation. Stop the boiler is produced after the cessation of the shedding of the powder from the walls of the device.
Use of freeze-drying according to the operating instructions of the installation that implements this process.
The finished product with a mass fraction of moisture not more than 10% is directed to filling and packaging.
Packaged products according to GOST 7630-96 in packages made of polymer materials OST 15-390-2005 maximum mass of 1.0 kg, followed by packing in boxes made of corrugated cardboard on the EAST 15-409-2002, GOST 13511-2006, GOST 13516-86 maximum mass of 20.0 kg Boxes glued adhesive tape on the paper base according to GOST 18251-87 or plastic tape with an adhesive layer according to GOST 20477-86.
Packaging products in bags made from polymer materials made in accordance with packing Instruction food fish products in bags and liners from plastic materials".
Containers and packaging materials used for packaging must be clean, solid, dry, free from foreign smell and made of materials permitted by the authorized bodies for contact with this type of product.
Collagen is stored at a temperature not above 20°C and relative to the air humidity not more than 80%, protecting from the sun's rays. The shelf life of the products with the manufacturing date is 24 months.
The proposed method of producing collagen economical, does not require the use of reagents. The final product has a neutral reaction and purified from salts and other impurities, which determines the possibility of its widespread use.
The liquid fraction obtained at the end of phase separation of the extract used for making drinks, preferably isotonic, for supplementing mineral-salt balance of the body of people working in difficult conditions or athletes, for example, mixing it with clean drinking water, sweeteners, poikilitically etc. ingredients.
A method of producing collagen from biological material, comprising the grinding of raw materials, liquid handling biological material with obtaining collagen containing substance that is separated into sludge and a liquid fraction, wherein the biological material used Medusa, preferably Rhopilema, preferably its dome, which is preferably crushed to 1-2 mm, to obtain collagen containing substance thus prepared material is mixed with potable water at a ratio by mass of the raw material to water as 1:2 and extracted at a temperature of preferably 15-18°C in t is the within 6-12 hours with periodic mixing, then the extract obtained is separated into a liquid fraction and a residue containing collagen, which is then dehydrated to the mass of moisture in it no more than 10%, then Packed and Packed.
SUBSTANCE: invention relates to biotechnology and can be used to produce biologically active collagen peptides from marine hydrobionts. Patiria pectinifera starfish is dehydrated with 96% ethyl alcohol and then demineralised with 1-2 N solution of an inorganic acid with ratio of the raw material to inorganic acid of 1:(3-5) for 1-3 days. The demineralised raw material is washed from traces of acid and water-soluble impurities with distilled water, after which the material is hydrolysed with an alkali solution with ratio of raw material to the alkali solution of 1:(3-5) in order to remove non-collagen proteins and washed with distilled water at temperature of 2-4°C. The obtained starfish collagen shells are homogenised. The homogenate is diluted with distilled water; pH of the suspension is brought to a value equal to 8.0-8.5 with an alkali solution and hydrolysed with 1% collagenase solution with ratio of homogenate to enzyme of (100-200):1 and temperature of 30-40°C for 3-5 hours, pH 8.5-7.0. The enzyme is inactivated at 80-90°C for 10-15 minutes. The hydrolysate solution is filtered to remove non-hydrolysed collagen, subjected to ultra-filtration through a 30 kD membrane filter to remove the inactivated enzyme; the end product, having antitumour, anticoagulant, wound-healing, anti-inflammatory, antioxidant activity, capacity to inhibit collagenase and angiotensin converting enzyme and which is a complex of collagen peptides with a high-molecular weight component weighing 22-23 kD, is concentrated in a vacuum and lyophilised.
EFFECT: obtaining a product, having antitumour, anticoagulant, wound-healing, anti-inflammatory and antioxidant activity.
2 cl, 7 tbl, 2 ex
SUBSTANCE: peptides consist of four amino-acid residues that are used for stimulation of collagen production with fibroblast.
EFFECT: invention allows effective stimulation of collagenoses in fibroblast cells.
11 cl, 3 dwg, 7 tbl, 6 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention discloses a medicine for treating a patient with rheumatoid arthritis on the basis of an antibody or a fragment thereof that bind the Extra Domain-A (ED-A) isoform of fibronectin. The antibody or fragment thereof contain the domain VH comprising a frame and a number of complementary-determining regions HCDR 1 -3, and the domain VL comprising a frame and a number of complementary-determining regions LCDR 1-3 with the antibody or fragment thereof being conjugated with IL-10. The invention discloses a medicine on the basis of the above antibody or fragment thereof for delivering a molecule conjugated with the antibody or fragment thereof wherein the molecule represents IL-10 to a newly formed vascular tree of the rheumatoid arthritis regions in a patient. There are disclosed method of treating rheumatoid arthritis with using the medicines under the invention, method of delivering IL-10 conjugated with the antibody or fragment thereof to the newly formed vascular tree of the rheumatoid arthritis regions in the patient, as well as a conjugate of the antibody or fragment thereof binding ED-A of fibronectin to interleukine-10. The antibody or fragment thereof may be represented by a diabody, contain one-chain Fv, present a small immunoprotein (SIP).
EFFECT: invention provides the successful treatment of rheumatoid arthritis ensured by the selective delivery of biologically active compounds conjugated on the antibody to the disease locations.
35 cl, 8 dwg, 2 tbl
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to medicine. The invention aims at creating biodegraded implants of the regenerated silk fibroin Bombyx mori. A method for preparing the biodegraded composite matrix of the regenerated silk fibroin Bombyx mori involves the stages: - dissolving the regenerated fibroin in a buffer containing CaCl2, C2H5OH, H2O in molar ratio 1:2:8. That is followed by dialysis of the prepared solution to the fibroin concentration min. 20 mg/ml. The fibroin solution of the concentration min. 20 mg/ml is mixed with dimethyl sulphoxide (DMSO). Gelatin or polylysine is added to the prepared mixture. The prepared mixture is frozen and thawed with an organic solvent added; what is also presented is the use of the specified biodegraded composite matrix as a base for the biodegraded implant.
EFFECT: group of inventions enables improving the biocompatibility, increasing the adhesive properties of matrix and cell proliferation in a mammalian body.
2 cl, 3 dwg, 3 ex, 1 tbl
SUBSTANCE: invention refers to the area of biotechnology, namely to production of short peptides - stimulators of production of extracellular matrix protein in the skin, and can be used in the medicine. The peptides produced consist of four amino-acid residues that can be used separately or in combination in the method of stimulation of collagen production using fibroblast.
EFFECT: invention provides for effective collagenesis stimulation in fibroblast cells.
25 cl, 3 dwg, 7 tbl, 6 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to medicine and concerns agents and methods based on using fibronectin domain EDA. Substance of the inventions involves using a polypeptide comprising an amino acid sequence of the fibronectin domain EDA, or its TLR4-binding fragment, or a version of specified domain; for preparing a pharmaceutical composition stimulating an antigen- specific immune response.
EFFECT: improved stimulant property.
36 cl, 3 ex, 11 dwg
FIELD: textile, paper.
SUBSTANCE: when boiling collagen, linear dimensions of leather tissue are measured before and after collagen boiling. The structure-to-structure distance is determined using difference of the sample thickness after boiling and the rated thickness of the sample before boiling, which is produced as a product of the sample thickness before boiling and a coefficient of layers number defined as a quotient from division of a lengthy sample length into the length of the sample after boiling. Invention makes it possible to realise the specified method objective.
EFFECT: method improvement.
4 ex, 3 tbl
SUBSTANCE: invention relates to low-molecular derivatives of peptides which are used for preparing a pharmaceutical agent which inhibits laminin/nidogen reaction.
EFFECT: increased effectiveness of compounds.
2 cl, 12 dwg, 2 tbl, 30 ex
FIELD: biotechnology, medicine.
SUBSTANCE: cells are contacted with protein containing fibronectin EDb-domen in presence of one or more chemical substances. As check experiment reaction between the same cells and abovementioned protein in absence of said substances. Expression of certain protein or presence of nucleic acid encoding the same makes it possible to select compounds bonding to fibronectin EDb-domen.
EFFECT: new biotechnological method.
1 tbl, 3 ex
FIELD: peptides, pharmacy.
SUBSTANCE: invention relates to low-molecular derivatives of peptides that are able to act as inhibitors in interaction between laminine and nidogen (interactions laminine/nidogen). Also, invention relates to a method for their preparing, pharmaceutical composition prepared on thereof and their using for preparing pharmaceutical agents, and for identification of inhibitors in interaction laminine/nidogen.
EFFECT: valuable properties of peptides.
5 cl, 12 dwg
SUBSTANCE: invention relates to a method for chemical conversion of a peptide chain into a peptide thioether. A -C(=X)-R1 group is incorporated into a thiol group of a cysteine residue and the obtained peptide then reacts in an organic solvent with a compound having a substituted group of formula: NH-C(=Y)NHR3, and a -NH-C(=Y)NHR3 group binds in an addition reaction with the carboxyl group of the peptide bond at the N-terminal side of the cysteine residue, through which the peptide bond is broken and the peptide moiety at the C-terminal end is cut off. When the obtained peptide chain, having a -NH-C(=Y)NHR3 group, reacts with thiol in a buffer solution, a thiol exchange reaction occurs, specifically the thiol group of the thiol compound binds with the carbon of the carbonyl to which the -NH-C(=Y)NHR3 was bound, thereby removing the -NH-C(=Y)NHR3 group.
EFFECT: achieving conversion to peptide thioether.
14 cl, 4 ex
SUBSTANCE: invention relates to a method of production of casein calcium chloride of technical casein by precipitation, and can be used in microbiological studies for production of components of storing media of cultures of microorganisms, and also production of calcium co-precipitates for food industry.
EFFECT: improvement of the method.
2 cl, 1 tbl, 5 ex
SUBSTANCE: disclosed is a method of purifying a compound of formula 1 which includes the following steps: (1) adding a raw compound 1 to a macroporous adsorption resin, (2) washing the macroporous adsorption resin with an aqueous solution, an organic solvent or a mixed solution of an organic solvent and water, (3) elution using the aqueous solution, organic solvent or mixed solution of an organic solvent and water.
EFFECT: improved method.
9 cl, 7 dwg, 12 ex
SUBSTANCE: invention relates to a method of purifying daptomycin, which includes steps a) loading partially purified daptomycin into an anion-exchange chromatographic column and subsequent purification steps b) and c) in reversed-phase chromatographic columns, where the elution buffer at step a) is a monovalent salt solution and the elution buffer at step b) and c) is an aqueous alcohol.
EFFECT: improved method.
16 cl, 2 ex
SUBSTANCE: invention refers to medicine and veterinary science and concerns a method for producing the purified antigen of Dirofilaria immitis. The presented method involves mechanical homogenisation, centrifugation of a homogenate, collection of a supernatant to be used as an antigen; the homogenisation involves a 2-cm head end of a mature female of Dirofilaria immitis placed in an aqueous solution of saccharose 0.25 M in a ratio of 1:3, frozen at a temperature of -18°C, that is followed by mechanical homogenisation and protein extraction in the aqueous solution of saccharose 0.25 M at 4°C for 12 hours, wherein 3 cycles of five 30-second ultrasonic homogenisations of the supernatant is performed at 70 kHz every 30 seconds at 0°C; the supernatant prepared after ultrasonic homogenisation is dissolved in cooled acetone at a temperature of 0°C in a ratio of 1:20 with exposition for 1 hour at a temperature of 4°C.
EFFECT: presented invention enables producing the high-sensitivity and specificity antigen and can be used in diagnosis of dirofilariasis in humans and animals.
2 tbl, 1 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: present invention refers to biotechnology, more specifically to allergen modifications to reduce an allergenic capacity thereof, and may be used in medicine. A modified allergen is prepared by sequential modification of all primary amino groups or a portion thereof of lysine and arginine residues of an allergen molecule with using potassium cyanate and phenylglyoxal and used as an ingredient of a pharmaceutical composition for treating allergy.
EFFECT: invention provides preparing the modified allergen possessing the reduced allergenic capacity as compared to a respective native allergenic material and allergoids prepared by modification by either cyanate, or phenylglyoxal.
9 cl, 12 dwg, 4 ex
SUBSTANCE: invention relates to biotechnology and represents a method of obtaining a preparation of an antibody or its antigen-binding site with reduced content of host cell proteins (HCP) from a sample mixture. The claimed invention can be used to obtain the antibody preparation. The method includes bringing pH to values 3-4 to form an initial separated sample. After that, pH of the initial separated sample is brought to values 6-8. The initial separated sample is brought in contact with resin for protein A-based affinity chromatography. The resin for affinity chromatography is washed out with a buffer solution. Sampling is carried out after affinity chromatography. The sample after affinity chromatography is brought in contact with an ion-exchange resin, after ion-exchange sampling is carried out. After the ion exchange sample is brought in contact with the resin for hydrophobic interaction chromatography (HIC) with sampling after HIC.
EFFECT: invention makes it possible to obtain the preparation of the antibody or its antigen-binding site with reduced HCP content.
25 cl, 12 dwg, 8 tbl, 4 ex
SUBSTANCE: group of inventions relates to the field of biotechnology. Claimed is a method of purification of a factor, contributing to wound healing, which represents a hepatocyte growth factor (HGF). All stages of purification are carried out in the presence of antithrombin III (AT-III). In accordance with the claimed method carried out are: defrosting of the frozen HGF-containing source and removal of sediment from the defrosted source. After that, the obtained solution, which contains a supernatant and AT-III, is brought in contact with a carrier for affinity chromatography on an immobilised heparin. Then, the solution is separated from the carrier for affinity chromatography. The carrier is brought in contact with a desorption buffer with ionic strength sufficient for HGF desorption. The desorption buffer, containing HGF, AT-III and histidine-rich glycoprotein (HRGP) is collected. Also claimed are wound-healing compositions, which contain HGF, AT-III and/or HRGP, purified by the claimed method.
EFFECT: inventions make it possible to increase step-by-step output of the hepatocyte growth factor, with the hepatocyte growth factor being concentrated in eluate in the presence of AT-III.
26 cl, 2 tbl, 6 ex
SUBSTANCE: invention relates to a method of producing glycoprotein which is uniform in terms of functions derived from a sugar chain (e.g., blood half-life) and physiological activity units, i.e., a glycoprotein, having uniform amino acid sequence, sugar chain structure and higher-order structure, physiological activity, a method for screening for glycoprotein and a method of producing a mixture of glycoproteins.
EFFECT: high efficiency of the method.
6 cl, 37 dwg, 4 tbl, 6 ex
SUBSTANCE: nucleotide sequences are formed, encoding the hybrid proteins EPO-TR 1.6, EPO-TR 4 and EPO-TR 6. Protein EPO-TR 1.6 is recombinant human erythropoietin fused with a fragment TR 1.6 of the human protein MUC1. Protein EPO-TR 4 is recombinant human erythropoietin fused with a fragment TR 4 of the human protein MUC1. The hybrid protein EPO-TR 6 is recombinant human erythropoietin fused with a fragment TR 6 of the human protein MUC1. Hybrid proteins are produced by the roller cultivation in the suitable conditions the modified mammalian cell line CHO containing a nucleotide sequence encoding the protein with subsequent isolation of the hybrid protein from the culture fluid.
EFFECT: invention enables to produce the hybrid recombinant human erythropoietin having the prolonged action.
4 cl, 4 dwg, 7 tbl, 9 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to the pharmaceutical industry, namely to a biologically active complex for organogenesis. The biologically active complex for organogenesis, is characterised by the fact, that a three-dimensional multi-component structure is organised by a three dimensional layer-by-layer replication of a native tissue of a human or animal structure, for the regeneration of which it is applied, for this purpose biopsy of the recipient's tissue is carried out, and in case of the absence of the tissue section with a need of its substitution an amino-acid composition of a protein component is determined by confocal microscopy, scanning and transmission electronic microscopy, a supramolecular structure is identified by a protein analyser, the composition of glycoseaminoglycanes and lipids is determined by the method of atomic-absorption analysis, then all basic protein components of the complex are synthesised by an autosynthesiser of proteins, glycoseaminoglycanes and lipids are selected qualitatively and quantitatively in ratios, maximally close to the ones obtained in biopsy analysis, then all the components are applied on a polymer substrate layer-by-layer, with the formation due to aggregation of the structure, similar to the recipient's own tissue, stem allogenic or autologous cells, covered by the following layer of proteins, lipids and glycoseaminoglycanes, are placed on each layer, the obtained complex is incubated and transplanted, closing the defect of the recipient's tissue.
EFFECT: complex described above makes it possible to reduce the period of rehabilitation due to the absence of a tissue, cellular and immune response of the recipient and, accordingly, fibrous changes in the area of the complex implantation.