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WO2004110484A1 - Procede de production d'un vaccin vivant de culture contre le virus de la grippe - Google Patents

Procede de production d'un vaccin vivant de culture contre le virus de la grippe Download PDF

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WO2004110484A1
WO2004110484A1 PCT/RU2003/000263 RU0300263W WO2004110484A1 WO 2004110484 A1 WO2004110484 A1 WO 2004110484A1 RU 0300263 W RU0300263 W RU 0300263W WO 2004110484 A1 WO2004110484 A1 WO 2004110484A1
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vaccine
influenza virus
cells
strains
influenza
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Lev Stepanovich Sandakhchiev
Elena Avgustovna Nechaeva
Nikolai Anatolyevich Varaksin
Tatyana Gennadyevna Ryabicheva
Natalya Alekseevna Mazurkova
Stanislav Georgievich Markushin
Yury Zakharovich Gendon
Irina Borisovna Koptyaeva
Irina Ivanovna Akopova
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Gosudarstvenny Nauchny Tsentr Virusologii I Biotekhnologii 'vektor'
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Gosudarstvenny Nauchny Tsentr Virusologii I Biotekhnologii 'vektor'
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Priority to PCT/RU2003/000263 priority Critical patent/WO2004110484A1/fr
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    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
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    • A61K39/00—Medicinal preparations containing antigens or antibodies
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    • A61K39/145—Orthomyxoviridae, e.g. influenza virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
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    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
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    • A61K2039/70—Multivalent vaccine
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    • C12N2760/16011—Orthomyxoviridae
    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
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    • C12N2760/00011—Details
    • C12N2760/16011—Orthomyxoviridae
    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
    • C12N2760/16151—Methods of production or purification of viral material
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    • C12N2760/16011—Orthomyxoviridae
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    • C12N2760/16111—Influenzavirus A, i.e. influenza A virus
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    • C12N2760/16011—Orthomyxoviridae
    • C12N2760/16211—Influenzavirus B, i.e. influenza B virus
    • C12N2760/16234—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • the invention relates to a technology for producing live influenza vaccine and can be used in the pharmaceutical industry for the production of vaccines and therapeutic drugs.
  • inactivated, subunit forms of influenza vaccines prepared on chicken embryos are produced and used to protect the public against influenza, which, when properly applied, can protect 80% of those vaccinated.
  • the high cost of such drugs, as well as the complexity of the production and use of inactivated vaccines make it difficult to use them for mass prevention of influenza.
  • inactivated vaccines form a higher level of general humoral immunity, these drugs are practically not able to induce local secretory immunity.
  • the effectiveness of an inactivated influenza vaccine decreases dramatically even with small changes in the antigenic structure of the epidemic strain compared with the vaccine.
  • a live vaccine In Russia, a live vaccine is widely used that stimulates the development of asymptomatic infection with the formation of humoral, secretory and cellular immunity with intranasal administration and immunological memory.
  • Sufficiently effective live vaccines have certain advantages over killed vaccines in terms of cost-effectiveness, ease of production and use, as well as a greater breadth of immunological effect. These benefits are associated with increased attention recently to the development of live influenza vaccine in the United States.
  • Currently used influenza vaccines need to be adjusted annually, taking into account the constantly occurring changes in the antigenic composition of the surface glycoproteins of the virus. Every year, an unpredictable situation arises when in a very short period of time it is necessary to produce a sufficient amount of vaccine that is adequately effective against strains that are most likely to cause an influenza epidemic.
  • Live influenza vaccine induces a wide range of antibodies that can neutralize variants with altered antigenic specificity of hemagglutinin.
  • a live vaccine is more economical because requires about 10 times less chicken embryos for production.
  • the use of cell culture in the production of vaccines makes the vaccine even more economical, and the use of a serum-free culture medium completely eliminates the allergic reactions of eggs vaccinated on chicken proteins.
  • a known method of producing an inactivated viral vaccine including culturing influenza virus strains in chicken embryos for 72-96 hours, incubating the embryos overnight in a refrigerator, collecting allantoic fluid, inactivating the virus, purification by filtration and / or centrifugation (Receptors For Ipastivative Ipfluepza Vassipe, World Independent Organization Testing Series, 384 (1966).
  • the disadvantages of this method are the content in the vaccine of certain amounts of chicken embryo proteins that cause adverse reactions in individuals allergic to these proteins; a change in the antigenic structure of the virus during cultivation in chicken embryos due to the loss of certain antigenic domains; the possibility of contamination with extraneous viruses; the need (one-time) for a large number of chicken embryos during vaccine production.
  • each chicken embryo is a unique biological system and thus suggests a certain instability of the resulting vaccine preparation.
  • Another method for producing a live viral vaccine is known.
  • Vaccine influenza allantoic live intranasal dry for adults including infection of chicken embryos with vaccine strains of the influenza virus, collection of virus-containing allantoic fluid, introduction of meat peptone, filling into ampoules, lyophilization and sealing.
  • the finished product is a dry porous mass with the smell of peptone.
  • the resulting vaccine does not contain chicken protein impurities.
  • this method has several disadvantages: a primary cell culture is used that does not have standard properties and is not sufficiently characterized; contamination of cell culture and vaccine with extraneous viruses, mycoplasmas, prions is possible; cell culture of chicken embryos is used, therefore, it is possible to change the antigenic structure of strains of the influenza virus, which occurs during cultivation on chicken embryos.
  • animal products are used (cattle blood serum, trypsin, gelatose or human serum albumin as a stabilizer), which can lead to allergenization of immunized , as well as a possible infection with extraneous viruses, mycoplasmas, prions contained in animal products.
  • the closest analogue is a method for producing inactivated and live influenza vaccines (US, 6344354, Bl, 02/05/2002), including cultivation of highly productive influenza virus strains on certified transplantable cell lines (Vero, MDCK) in nutrient medium MEM Needle with the addition of 5 -10% fetal serum and trypsin in an amount (0.05-1.0 ⁇ g / ml), washing the cells after the formation of a monolayer, introducing bovine serum albumin, collecting virus-containing liquid, purification by centrifugation or chromatography and subsequent iv denie chemotherapeutic components, adjuvants or polymers (polyethylene glycol, polypropylene glycol).
  • the disadvantages of this prototype method is a monolayer cultivation of cells in culture bottles, which cannot ensure the production of large volumes of virus-containing fluid; the use of MEM Needle for the cultivation of cells of the nutrient medium with the addition of 5-10% fetal serum, as well as the introduction of bovine serum albumin virus at the stage of production, leads to the introduction of components of animal feed that can be a source of extraneous viruses, mycoplasmas, prions, and cause additional allergization of the immunized organism.
  • the technical result of the proposed method is the production on an industrial scale of a live cultural trivalent influenza vaccine based on cold-adapted vaccine strains of influenza A and B virus for intranasal and parenteral administration with a lower content of extraneous protein components of animal origin, which reduces the possibility of allergic reactions.
  • cold-adapted vaccines are used as viral strains sorts of strains of influenza viruses, as a nutrient medium in the cultivation of MDCK cells and strains of influenza viruses on these cells use serum-free culture medium with the addition of soy hydrolyzate in a concentration of 0.01- 10% and a proteolytic enzyme in an amount of 0.25-50.0 ⁇ g / ml before the inoculation dose of influenza virus strains, and the cultivation of MDCK cell cultures and cold-adapted influenza virus strains in the specified cell culture is carried out in suspension on microcarriers introduced into serum-free nutrient medium at a concentration of 1-6 g
  • a culture of MDCK cells As a culture of MDCK cells, a culture stored at liquid nitrogen temperature in the form of a seed and working banks and certified for the absence of extraneous viruses, the absence of oncogenic potentials and species identity is used.
  • vaccine strains of type A / HZN2 are used; A / H1N1 and type B of this virus.
  • the seeding dose of transplanted MDCK cells is 100-600 thousand cells per 1 ml of medium, and the seeding dose of vaccine strains of influenza virus has a multiplicity of infection of 0.01-0.0001 EIDso / cell.
  • Papain is a proteolytic enzyme of plant origin as a proteolytic enzyme, and Cytodex 1 is used as a microcarrier.
  • the virus-containing fluid is collected after the specific activity of the influenza virus is at least 8.0 Ig EID 5 o / ml, and the ballast impurities are removed from the viral substance by filtration .
  • sucrose and peptone from soybeans are used in the final concentrations of (5-10) and (3-8) May. % respectively.
  • Drying the vaccine is carried out by freezing it for 18 hours at a temperature not exceeding minus 4O 0 C followed by lyophilization for 48 hours, moreover, before sealing, the ampoules with the viral vaccine are filled with an inert gas, in particular argon.
  • Vaccine strains of live influenza vaccine are reassortants obtained in Russia on the basis of cold-adapted mutant donors of attenuation A / Leningrad / 134/47/57 (H2N2) and B / CCCP / 60/69. These donors are characterized by cold adaptation, i.e. the ability to reproduce at low temperatures, temperature sensitivity and harmlessness for laboratory animals and humans.
  • a necessary condition for the safety and genetic stability of vaccine strains is considered to be the presence of 5-6 mutant genes encoding the internal proteins of the attenuation donor in their genome, while the hemagglutinin (HA) and neuraminidase (NA) genes always come from the current epidemic virus.
  • Strain A / 47 / Chandon / 93/1 (H1N1) was obtained by crossing the cold-adapted master strain A / Leningrad / 134/47/57 (H2N2) with the epidemic strain A / Chandon / 93/1 (H1N1).
  • influenza virus strain A / 47 / Johannesbyrg / 94/2 was obtained by crossing the cold-adapted master strain A / Leningrad / 134/47/57 (H2N2) with an epidemic strain
  • Strain B / 60 / Peterbyg / 95/20 was obtained by crossing a cold-adapted master strain B / CCCP / 60/69 with the epidemic strain B / Peterbyg / 95/20.
  • Vaccine strains of the influenza virus are stored at the State Research Institute for Standardization and Control of Medical Biological Products (GISK) named after L.A. Tarasevich, Moscow.
  • a significant advantage of transplanted cells is their stability, standardization, the possibility of obtaining a large number of producer cells in a fairly short time, long-term use of cells, and the relatively low cost of vaccine preparations.
  • WHO-certified cell lines are free of contaminating agents, while chicken embryos often contain retroviruses and some other agents.
  • the hemagglutinin of human influenza virus strains isolated and cultured in chicken embryos may lose one of the antigenic domains, while the isolation and cultivation of the influenza virus on the MDCK cell line preserves all antigenic domains, and the hemagglutinin of such isolates is antigenically completely identical to hemagglutinin influenza virus that multiplies in the human body.
  • vaccines prepared on MDCK or Vero cells better protect vaccinated animals from infection with virulent influenza virus strains and also induce more cross-reactive antibodies than vaccines obtained on chicken embryos that stimulate antibodies specific for to the homologous antigen (Alumova L, Codihalli S., Govorkova E. et al. Immune aproprotect effisasu ip miise odfluepza virus Vassipes grpwa ipma mma).
  • MDCK cell line derived from dog kidney cells mainly the MDCK cell line derived from dog kidney cells and the Vero cell line derived from African green monkey kidney cells were investigated.
  • comparative experiments (Mertep O., Managuerra J., Hannoun C. et al. Rupodtiop of ipfluepza virus ip sepulcultures for preserving reproduct.
  • Novel Str. Rlepspres N.-Y., 1996, pp 141-150
  • it was shown that MDCK cells produce the same number of cells 2 times faster than Vero cells and 7 times faster than BHK cells.
  • Papain is a plant proteolytic enzyme of the hydrolase class; it exhibits broad substrate specificity and is produced by the company
  • Papain provides in a nutrient medium at the indicated concentrations the guaranteed penetration of the influenza virus into MDCK cells during their infection with the indicated virus.
  • Microcarriers provide at the indicated concentrations higher production of cells and virus and a higher yield of virus-containing material. In the process of experimental studies, it was precisely the Sigma Cytodex 1 microcarriers that were selected that were 1 mm in size, on the surface of which stable monolayers of MDCK cells are formed, which are capable of producing the influenza virus without losing their physiological functions.
  • the serum-free nutrient medium MDSS2 is manufactured by Ahsevir, France. Embodiments of the invention
  • Example JVa Certification of MDCK cell culture. For work using a culture of MDCK cells from the seed and working banks. Certification of cell culture MDCK carried out in accordance with the requirements of the WHO for transplantable cell lines.
  • the cytomorphology of culture in native and fixed preparations in a light microscope was studied. It was shown that the MDCK cell culture consists of epithelial-like cells. The nuclei are rounded with chromatin grit. Nuclei rounded, from one to several in the nucleus.
  • the cytoplasm is granular. There are giant and multinucleated cells, the number of which increases during cultivation. Cells form a monolayer on the 2nd day of cultivation, at the same time, a peak in mitotic activity is noted, pathological mitoses are not observed.
  • Mycoplasma was not detected by direct inoculation of the culture fluid of the seed and working banks of MDCK cells on selective nutrient media and by DNA staining with intercalating fluorochromes.
  • two pairs of primers were used (GPO-I and MGSO-for the first cycle; GPO-2 and MGSO-for the second cycle), which make it possible to identify all known types of mycoplasmas.
  • Mycoplasma was not detected by PCR analysis.
  • a control was conducted to detect foreign viruses in cell culture, chicken embryos and animals.
  • the state of the monolayer of intact cells of the inoculating and working banks was studied for 14 days. No cytopathic effect (CPP) was found.
  • samples of the culture fluid were examined in a hemagglutination reaction, and the monolayer in the hemadsor sion reaction with guinea pig erythrocytes. The reactions are negative.
  • Culture fluid samples were studied in three types of cell cultures: MDCK (homologous), L-68 (diploid culture), Vero for 14 days. No CPC detected.
  • the studied material in the amount of 10 6 cells was introduced into the yolk sac, allantoic cavity and the chorionallantoic membrane chicken embryos (10 embryos per point), the embryos were incubated for 7-10 days, after which the viability of the embryos was determined and a hemagglutination reaction with allantoic fluid was established. No foreign viruses were detected in the seed and working banks of the MDCK line.
  • the studied material was intramuscularly administered to sucker mice at the age of 24 hours (28 heads of sucker mice, 10 7 cells per group), adult mice (10 goals, 10 7 cells per group of mice), and guinea pigs (5 animals, 10 7 cells per group), rabbits (5 animals, 10 7 cells per animal group), the animals were monitored for 28 days, after which autonasia and examination of the internal organs were performed. All animals in the experiment were alive, no changes in internal organs were noted. No extraneous viruses were found in the seed and working banks of MDCK cells.
  • the oncogenic potencies of the cells of the seed and working banks were determined in the ip vivo system.
  • the method of heterotransplantation of cells to immunosuppressive animals was used in comparative experiments with a reference HeIa cell line that causes progressive tumor growth. The animals were observed for 21 days. In this case, all animals were alive, with a macroscopic examination for the presence of a tumor in the lymph nodes, liver, kidneys, lungs and at the injection site, pathological changes were not detected.
  • monitoring in the ip vivo system it was established that the MDCK cells of the seed and working banks do not possess oncogenic potentials.
  • the cells of the seed and working banks of MDCK have a fairly stable karyotype, characterized by clear morphological characteristics. An analysis of the electrophoretic mobility of the G-6-FDG isoenzyme was carried out.
  • the MDCK cells of the seed and worker banks have G-6-FDG characteristic of dog cells.
  • Example JVa Preparation of a living culture influenza vaccine (VHF) substance.
  • a culture vessel of a 1 or 10 liter fermenter is filled with serum-free culture medium.
  • a certified MDCK cell culture (in accordance with Example 1) is taken from a collection bank of cell cultures of the State Scientific Center of the WB Bektop and brought into the indicated culture vessel at the rate of 100-600 thousand cells per ml of culture medium.
  • the microcarrier Cytodex I is introduced into the culture vessel at a concentration of 1-6 g / l.
  • Cell deposition on a microcarrier is carried out for 12 hours at a temperature of 35-37 0 C (the filling volume of the culture vessel is 25-30%).
  • Cell planting mode 5 min stirring, 10 min break.
  • the specified cultivation parameters the stirring device rotational speed is 50-80 rpm, surface aeration with a blowing rate from 0 to 10 l / h.
  • the pH is maintained between 6.9 and 7.2 with a dosed supply of a 7.5% sodium carbonate solution.
  • the cultivation temperature is 35-37 0 C.
  • Perfusion of the nutrient medium begins at a cell concentration of 600-900 thousand / ml. The perfusion rate depends on the concentration of cells.
  • the cell cultivation time is 96-144 hours. After the accumulation of cells, 50% of the medium is removed, one of the vaccine cold-adapted reassortants of the virus is introduced into the suspension 63
  • influenza for example, A / 47 / Johannesbyrg / 94/2 (HZN2), with a multiplicity of infection of 0.01-0.0001 EID 50 / cell, and supporting a serum-free nutrient medium containing the papain proteolytic enzyme in an amount of 0.25-50, 0 ⁇ g / ml or trypsin in an amount of 2.0-20.0 ⁇ g / ml and soy hydrolyzate in a concentration of 0.01-10%.
  • the virus is cultivated at a temperature of (34 + I) 0 C.
  • a virus-containing liquid is collected with a specific activity of at least 8.0 Ig EID 50 / ml, which is released by filtration from cell detritus .
  • a sterile stabilizer solution (sucrose and peptone from soybeans at a final concentration of 10% and 5%, respectively) is added to the virus-containing suspension to obtain a liquid semi-finished product of strain A / 47 / Johannesbyg / 94/2 (HZN2) of the live influenza vaccine (GHV).
  • liquid semi-finished vaccine strains of cold-adapted reassortants of the influenza virus A / 47 / Chandon / 93 / l (HlNl) and B / 60 / Peterbyg / 95/20 are produced in parallel or sequentially.
  • EID is an embryonic infectious dose.
  • liquid semi-finished products obtained from the virus-containing liquid of vaccine strains A / 47 / Johannesbyrg / 94/2 (HZN2), A / 47 / Chandon / 93 / l (HlNl) and B / 60 / Peterbyg / 95/20 influenza virus are combined in equal proportions.
  • a liquid semi-finished product of one of the above strains of the influenza virus is used.
  • the resulting liquid form of the vaccine is poured into ampoules of 0.6 ml each and frozen at a temperature not exceeding minus 4O 0 C for 18 hours.
  • the product is freeze-dried for 48 hours under sterile conditions. After drying, the ampoules are filled with an inert gas (argon) and sealed.
  • Example JVa 4 Determination of the specific activity of preparations of live influenza vaccine
  • the specific activity of the virus-containing material is determined in monovaccines to the information of the trivalent vaccine obtained simultaneously with the trivaccine, by titration on chicken embryos according to the method described in (Methodological instructions "Methods of control of medical immunobiological preparations administered to people.” MUK 4.1 / 4.2.588-96 )
  • the specific activity of the vaccine after lyophilization is not less than 6.0-6.5 Ig EID 5 o / O, 2 ml.
  • Example JVs 5 The study of the dynamics of reproduction of the influenza virus with different multiplicity of infection on the example of a strain
  • the MDCK cell culture is infected with strain A / 47 / Chandon / 93 / l (HlNl) of influenza virus with different multiplicity of infection (0.001;
  • Example JVs 6 The study of the stability of ts ⁇ 4 o and and ca 25 genetic traits
  • Example JNa 7 Study of the effect of the papain enzyme on the cultivation of cold-adapted vaccine strains of the influenza virus In order to replace the components of animal origin in the technology for producing live influenza vaccine with herbal preparations, studies were carried out on the use of the plant papain enzyme in the production of influenza virus strains. To do this, papain is added to the support medium used in the cultivation of influenza virus strains. The data are shown in table 3. Table 3
  • table 3 shows that at a concentration of papain in a support medium of 0.25-20.0 ⁇ g / ml, the titer of the influenza virus on 3-4 days of cultivation was 7.3-8.5 Ig EID 5 (/ ml, while in the control experiment (without introducing the enzyme into the supporting medium) the titer of the influenza virus was 6.5-6.7 Ig EID 5 o / ml, which confirms the effectiveness of the specified enzyme.

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  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention relève du domaine de la biotechnologie, et a trait à un procédé de production d'un vaccin vivant contre la grippe. Le procédé selon l'invention consiste à cultiver des cellules MDCK entrelacées dans un milieu nutritif, et à produire une substance contenant le virus en cultivant des souches vaccinales du virus de la grippe dans une culture de cellules MDCK, sur le même milieu nutritif. Les souches vaccinales utilisées sont des réassortis vaccinaux des souches des virus de la grippe résistants au froid. Le milieu nutritif, lors de la culture des cellules MDCK et des souches des virus de la grippe sur lesdites cellules, se présente sous la forme d'un milieu nutritif sans sérum auquel l'on a ajouté un hydrolysat de soja, dont la concentration est comprise entre 0,01 et 10 %, et un ferment protéolytique d'origine végétale, dont la quantité est comprise entre 0,25 et 50,0 mcg/ml, avant d'ajouter une dose d'inoculation des souches des virus de la grippe. La culture des cultures de cellules MDCK et des souches vaccinales du virus de la grippe dans ladite culture de cellules est réalisée sur des microporteurs en suspension, qui sont introduits dans le milieu nutritif sans sérum à une concentration de 1 à 6 g/l. On lave ensuite la suspension virale des impuretés indésirables, on ajoute un stabilisateur et on la sèche.
PCT/RU2003/000263 2003-06-18 2003-06-18 Procede de production d'un vaccin vivant de culture contre le virus de la grippe Ceased WO2004110484A1 (fr)

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PCT/RU2003/000263 WO2004110484A1 (fr) 2003-06-18 2003-06-18 Procede de production d'un vaccin vivant de culture contre le virus de la grippe

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PCT/RU2003/000263 WO2004110484A1 (fr) 2003-06-18 2003-06-18 Procede de production d'un vaccin vivant de culture contre le virus de la grippe

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JP2008525025A (ja) * 2004-12-23 2008-07-17 メッドイミューン バクシーンズ，インコーポレイティド ウイルス増殖用の非腫瘍形成性ｍｄｃｋ細胞株
EP1862537A4 (fr) * 2005-03-10 2009-01-21 Kyoritsu Seiyaku Corp Lignée cellulaire cultivable sans composant dérivé d'un animal, procédé destiné à l'établir, procédé de production de virus utilisant la lignee cellulaire et procede de production de vaccins
US8202726B2 (en) 2008-09-24 2012-06-19 Medimmune, Llc Methods for cultivating cells, propagating and purifying viruses
CN102660511A (zh) * 2012-05-17 2012-09-12 肇庆大华农生物药品有限公司 一种降低血清中胰酶抑制物的方法及其应用
US8357376B2 (en) 2006-09-15 2013-01-22 Memimmune, LLC Method of purifying influenza virus and removing MDCK cell DNA contaminants
CN107460156A (zh) * 2016-06-03 2017-12-12 北京大北农科技集团股份有限公司动物医学研究中心 无血清全悬浮mdck细胞株及其在生产流感病毒中的应用
RU2701953C1 (ru) * 2018-12-28 2019-10-02 Общество с ограниченной ответственностью "Нанолек" Способ получения поливалентной вакцины от гриппа
RU2701959C1 (ru) * 2018-12-28 2019-10-02 Общество с ограниченной ответственностью "Нанолек" Способ контроля получения продукта вакцина от гриппа
RU2706191C1 (ru) * 2018-12-28 2019-11-14 Общество с ограниченной ответственностью "Нанолек" Поливалентная вакцина против гриппа
RU2706544C1 (ru) * 2018-12-28 2019-11-19 Общество с ограниченной ответственностью "Нанолек" Концентрат вакцины от гриппа и способ его получения

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JP2012210225A (ja) * 2004-12-23 2012-11-01 Medimmune Llc ウイルス増殖用の非腫瘍形成性ｍｄｃｋ細胞株
US7670837B2 (en) 2004-12-23 2010-03-02 Medimmune, Llc Non-tumorigenic MDCK cell line for propagating viruses
US8119388B2 (en) 2004-12-23 2012-02-21 Medimmune, Llc Non-tumorigenic MDCK cell line for propagating viruses
US8748174B2 (en) 2004-12-23 2014-06-10 Medimmune, Llc Non-tumorigenic MDCK cell line for propagating viruses
JP2008525025A (ja) * 2004-12-23 2008-07-17 メッドイミューン バクシーンズ，インコーポレイティド ウイルス増殖用の非腫瘍形成性ｍｄｃｋ細胞株
EP1862537A4 (fr) * 2005-03-10 2009-01-21 Kyoritsu Seiyaku Corp Lignée cellulaire cultivable sans composant dérivé d'un animal, procédé destiné à l'établir, procédé de production de virus utilisant la lignee cellulaire et procede de production de vaccins
US7910366B2 (en) 2005-03-10 2011-03-22 Kyoritsu Seiyaku Corporation Cell strain capable of being cultured without ingredients derived from animals, method of producing the same, method of producing virus using the same, and method of producing vaccine
US8357376B2 (en) 2006-09-15 2013-01-22 Memimmune, LLC Method of purifying influenza virus and removing MDCK cell DNA contaminants
US8846032B2 (en) 2006-09-15 2014-09-30 Medimmune, Llc MDCK cell lines supporting viral growth to high titers and bioreactor process using the same
US8202726B2 (en) 2008-09-24 2012-06-19 Medimmune, Llc Methods for cultivating cells, propagating and purifying viruses
US9085753B2 (en) 2008-09-24 2015-07-21 Medimmune, Llc Methods for cultivating cells, propagating and purifying viruses
CN102660511A (zh) * 2012-05-17 2012-09-12 肇庆大华农生物药品有限公司 一种降低血清中胰酶抑制物的方法及其应用
CN102660511B (zh) * 2012-05-17 2013-08-21 肇庆大华农生物药品有限公司 一种降低血清中胰酶抑制物的方法及其应用
CN107460156A (zh) * 2016-06-03 2017-12-12 北京大北农科技集团股份有限公司动物医学研究中心 无血清全悬浮mdck细胞株及其在生产流感病毒中的应用
CN107460156B (zh) * 2016-06-03 2022-03-11 兆丰华生物科技(南京)有限公司北京生物医药科技中心 无血清全悬浮mdck细胞株及其在生产流感病毒中的应用
RU2701953C1 (ru) * 2018-12-28 2019-10-02 Общество с ограниченной ответственностью "Нанолек" Способ получения поливалентной вакцины от гриппа
RU2701959C1 (ru) * 2018-12-28 2019-10-02 Общество с ограниченной ответственностью "Нанолек" Способ контроля получения продукта вакцина от гриппа
RU2706191C1 (ru) * 2018-12-28 2019-11-14 Общество с ограниченной ответственностью "Нанолек" Поливалентная вакцина против гриппа
RU2706544C1 (ru) * 2018-12-28 2019-11-19 Общество с ограниченной ответственностью "Нанолек" Концентрат вакцины от гриппа и способ его получения

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