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Method of virus treatment by ultracentrifugation in sugar concentration gradient (versions) Treatment methods of inactivated virus or fragmented virus (versions) are proposed. Methods involve addition of virus preparation to sugar gradient and further centrifugation to obtain peak fraction. Then, peak fraction is extracted to obtain cleaned virus. With that, sugar gradient is obtained by continuous ultracentrifugation of the first and the second buffer sugar solution. Besides, the first buffer sugar solution includes the first physiological buffer, and the second buffer sugar solution includes the second physiological buffer that is the same or different from the first physiological buffer. As per one version, sugar concentration in the first buffer solution is equivalent to saccharose concentration of 35 to 50 wt %, and sugar concentration in the second buffer solution is equivalent to saccharose concentration of 50 to 65 wt %. Besides, the second buffer sugar solution has higher density in comparison to that of the first one. As per the other method version, density of the first buffer sugar solution is 1.15 kg/l to 1.23 kg/l, and density of the second one is 1.23 kg/l to 1.32 kg/l. |
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Method for preparative extraction of viruses of plants Homogenisation of tissues of an infected plant is performed in neutral buffer solution based on 2-[4-(2-hydroxyethyl)piperazin-1-il]ethanesulphonic acid (HEPES) in concentration of 0.02-0.03 M with addition of inhibitors of plant ferments of iodine-sodium acetate and phenilmethylsulfonyl fluoride. Saccharose is added. Obtained extract is clarified. Clarified extract is treated with 4 - 6% Triton X-100 to separate viral particles from cell components. Viral particles are separated from clarified extract by differential centrifugation or ultracentrifugation. |
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Method for producing preparation containing viral antigens and using preparation Invention refers to molecular biology, genetic engineering and virology. What is presented is a method for producing a preparation containing the viral antigens, involving a) cell inoculation with an infectious in the fluid, b) reproduction of the above virus in the above cells, c) collection of the above reproduced virus, d) inactivation of the above collected virus, and e) treatment of the above inactivated virus with a detergent to produce the preparation containing the viral antigens. |
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Method for influenza virus purification Invention refers to biotechnology and describes a method for preparing a purified concentrate of influenza virus; the method involves microfiltration of a virus-containing allantoic fluid in filter elements of the size of 0.1-0.2 mcm and further diafiltration in a phosphate buffer solution containing sodium chloride 0.25-0.5 M, EDTA or sodium citrate, concentration of influenza virus on membranes with a cuttoff threshold of 300-500 kDa and further diafiltration in a phosphate buffer solution containing sodium chloride 0.25-0.5 M, EDTA or sodium citrate, an anionic detergent 0.02-0.002%, and ultra-centrifugation in a density gradient of saccharose prepared in a phosphate buffer solution containing sodium chloride 0.25-0.5 M, EDTA or sodium citrate, and an anionic detergent 0.02-0.002%. |
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New method for recovery and viral load test in pancreatine sample Invention refers to biotechnology. The method for viral load recovery from a pancreatine sample provides: preparation of a pancreatine test sample, low-speed centrifugation, separation of a solid deposit, first ultracentrifugation, separation of a first target fraction, second ultracentrifugation and viral load recovery. The low-speed centrifugation is performed at relative centrifugal force 8000-15000 x g. The first ultracentrifugation follows at relative centrifugal force 50000-150000 x g, and relative centrifugal force for the second ultracentrifugation is 150000-300000 x g. |
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Method of cleaning intended to produce cleaned virus of vesicular stomatitis from cellular culture Method includes clarification of a cellular culture liquid by low-speed centrifugation at the speed of lower than 10000 rpm or with the help of a module of volume filtration, the pore size of which makes from 1.0 mcm to 4.5 mcm, to produce a vesicular stomatitis virus in a supernatant. The supernatant is filtered via a filter with holes of the diameter of 0.2-0.45 mcm, and the vesicular stomatitis virus is produced in the filtered solution. The filtered solution, which contains the vesicular stomatitis virus, is loaded to an anion-exchange membrane adsorber balanced with the first buffer salt solution. The first buffer salt solution has ion force from 100 mM to 400 mM. The vesicular stomatitis virus is eluted from the anion-exchange membrane adsorber with the second buffer salt solution. The vesicular stomatitis virus is eluted from the membrane adsorber, when the second buffer salt solution is added, in one stage. Concentration of salt in the single-stage elution makes from 500 mM to 750 mM. Eluted fractions containing the vesicular stomatitis virus are collected. The vesicular stomatitis virus is cleaned with running filtration along the flow direction (press filter) using the membrane, the transmission limit of which is from 300 kDa to 1000 kDa, and the concentrate of the vesicular stomatitis virus is produced. The concentrate containing the vesicular stomatitis virus is filtered via a filter with holes of the diameter of 0.2-0.22 mcm, and the vesicular stomatitis virus is produced in the filtered solution. The method of cleaning does not include use of human embryos. |
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Method for purifying mammalian and human recombinant adenoviruses Method involves collection of adenoviruses in a permissive cell culture of line HEK-293, collection of these cells, release of recombinant viruses due to cell destruction by re-frosting, and purification. It involves the use of the recombinant capside-modified adenovirus with modified capside protein pIX carrying a polysaccharide-binding domain specified in a group consisting of a cellulose-binding domain, a dextran-binding domain. The recombinant capside-modified adenoviruses are implanted on the polysaccharide carrier specified from a group consisting of a cellulose carrier for binding with the cellulose-binding domain, the dextran carrier - for binding with the dextran-binding domain. The mixture is incubated. It is followed by the process of purification; for this purpose the polysaccharide carrier with the adenoviruses is settled by centrifugation, while the prepared supernatant is removed. The settled polysaccharide carrier with the adenoviruses is washed in washing buffers, then the adenoviruses are eluted from the polysaccharide carrier. The prepared suspension is separated by centrifugation. The virus-containing supernatant is selected and stabilised. |
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What is described is a method for stabilising a Newcastle disease virus for storage in an aqueous solution for 6 months or more at temperature -30°C to -10°C. The method provides preparing the aqueous solution containing the Newcastle disease virus and saccharose or trehalose. The solution contains ten portions per million or less of each of reducing substances, antioxidants, sodium chloride, dextrane, mannitol, sorbitol, Tween, glutamate, polyethylene glycol, calcium chloride, phosphatidylcholine, glycine and phosphate; it has pH 7-9 and osmotic pressure 250-300 milliosmole. |
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Method of obtaining life culture vaccine against influenza virus Invention relates to field of medicine and deals with method of obtaining lice culture influenza vaccine. Essence of the invention includes method of obtaining virus-containing substance by cultivation of one of cold-adapted influenza virus reassortants with enoculation dose, with multiplicity of infection not lower than 0.0001 EID50/cell in MDCK cell culture on micro-carriers, which have concentration not less than 1 g/l, with application as micro-carrier material of porous polypropelene, in supporting serum-free nutritional medium, containing proteolytic enzyme in amount 0.25-50.0 mcg/ml, and stabilising additive, which includes sorbitol, or sucrose, or peptone from soya in concentration 0.5-4.0 wt%, collection of virus-containing liquid after cultivation is carried out at least 2 times when specific influenza virus activity before each collection of virus-containing liquid reaches at least 7.0 Ig EID50/ml, concentration and purification of virus substance from ballast admixtures, introduction into purified substance before drying of stabilising additives, with application as such of either proline, glycene, lactose, glutamine-acidic sodium, sucrose, gelatins in final concentration (1.5-5), (1.5-5), (1.5-10), (1.5-5), (5-30) and (1-10) wt % respectively, or sucrose, gelatose and soya peptone in final concentration (1-8), (1-8) and (1-8) wt % respectively, or sorbitol and gelatose in final concentration (3-8) and (3-8) wt % respectively. |
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Method of obtaining virion antigen of tick-borne encephalitis virus Invention relates to field of biotechnology, to method of obtaining virion antigen of tick-borne encephalitis (TBE) virus from antigen-containing material, which includes, mainly, antigen particulars of TBE virus smaller then full-size TBE virions. Claimed method includes centrifuging of antigen-containing material, which includes, mainly, antigen particles of TBE virus smaller than full-size TBE virions with angular acceleration not less than 15000 g. |
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Medium for cells cultivation that does not contain animal proteins Invention may be used for considerable reduction of variations in produce of recombinant proteins, which occur in process of cells cultivation with application of various batches of commercially available soy-bean hydrolysate. Medium is produced for cells cultivation, which does not contain animal proteins, by means of complementing of mediums, which do not contain animal proteins, with soy-bean hydrolysate and additionally with biogenic amine in the range of 1-18 mg/l. Produced medium is used to produce recombinant proteins by means of cultivation of according cells. |
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Invention refers to biotechnology and virology. Method of virus replication provides cultivation of permissive cells on an insoluble porous carrier, further infection thereof with virus to be replicated in the cells attached to the carrier. Said permissive cells are those of human or animal. The virus replicable in such cells functions as an infecting, and the insoluble porous carrier is polymer cryogel with connected pores of section 50-500 mcm. As a result, offered invention implemented enables the effect of eliminated undesired clogging of the carrier pores with the growing culture and cell debris, ensures greater universality of the method with respect to permissive cell types. Herewith more effective use of the whole carrier and culture medium with reduced risk of failed sterility and cost price of an end product, higher yield of the end virus. |
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Lung cells of cotton rats for cultivation of viruses Method is suggested for production of virus on lung cells of cotton rats, and also line of lung cells of cotton rats ATCC PTA-3930 for growth, reproduction and cultivation of viruses. |
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Device for cleaning virion structures of flaviviruses Device comprises series-arranged cathode electrode vessel, cathode accumulation chamber and anode accumulation chamber separated by a barrier, and an anode electrode vessel. The cathode and anode electrode vessels are separated by dialysis parchments from the cathode and anode accumulation chambers respectively. Between the dialysis parchment, separating the anode accumulation chamber from the anode electrode vessel, and the anode electrode vessel is a buffer chamber and an agarose gel unit with high degree of electro-endosmosis placed in series. |
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Method of virus concentration from liquid media Invention claims virus adsorption on metacrylic acid hydrogel of at least 2.0 mg/ml concentration, sorption at pH 5.0 and 4 to 22°C for 20 minutes to 24 hours, and further centrifugation of virus-sorbent complex and virus elution. |
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Method for large-scale production of viral antigen Method involves preparing culture of adhesive cells, their growing, infecting cells with virus and incubation of cells. The density of cells in biomass of cell culture grown before their fusion is increased by at least 1.3-fold as compared with value before and after infecting cell with virus. Method provides enhancing producing virus in small volume of the cellular culture. Invention can be used in medicine and veterinary science. |
Another patent 2513702.