WO2009013551A2 - Procédé de préparation d'un vaccin viral - Google Patents

Procédé de préparation d'un vaccin viral Download PDF

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Publication number
WO2009013551A2
WO2009013551A2 PCT/HR2008/000024 HR2008000024W WO2009013551A2 WO 2009013551 A2 WO2009013551 A2 WO 2009013551A2 HR 2008000024 W HR2008000024 W HR 2008000024W WO 2009013551 A2 WO2009013551 A2 WO 2009013551A2
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WIPO (PCT)
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egg
ndv
ibdv
virus
vaccine composition
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Ceased
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PCT/HR2008/000024
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WO2009013551A3 (fr
Inventor
Stanislav Cajavec
Neda Ergotic
Andrej Cizelj
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Veterina doo
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Veterina doo
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Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/76—Viruses; Subviral particles; Bacteriophages
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A61K39/155—Paramyxoviridae, e.g. parainfluenza virus
    • A61K39/17—Newcastle disease virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A61K39/295—Polyvalent viral antigens; Mixtures of viral and bacterial antigens
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • C12N7/02—Recovery or purification
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/525—Virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
    • A61K2039/552—Veterinary vaccine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/70—Multivalent vaccine
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2720/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsRNA viruses
    • C12N2720/00011—Details
    • C12N2720/10011—Birnaviridae
    • C12N2720/10034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/18011—Paramyxoviridae
    • C12N2760/18111—Avulavirus, e.g. Newcastle disease virus
    • C12N2760/18134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • the present invention relates to a process for the preparation of viral vaccines, in particular vaccines containing a combination of two or more viruses.
  • the process is particularly suitable for the preparation of vaccines against common avian diseases, especially Newcastle Disease and infectious bursal disease (Gumboro Disease).
  • Newcastle disease presents an exceptional risk for the poultry industry in almost all countries of the world, irrespective of whether it concerns intensive or extensive, poultry production. The global economic damage caused by this disease is enormous.
  • Newcastle disease virus which contains a non- segmented, single-chain RNA and belongs to the genus Paramyxovirus of the family Paramyxoviridae.
  • NDV Newcastle disease virus
  • PMV-I the group of avian paramyxoviruses
  • PMV-I the group of avian paramyxoviruses
  • Pathogenic NDV endangers all domestic fowl, although chickens and turkeys are most susceptible. Significant losses (mortality, drop in egg yield) can also occur in wild fowl farming management (Alexander, D. J. (2003): Newcastle Disease, Other Avian Paramyxoviruses, and Pneumovirus Infections. Pages 63-99. In: Diseases of Poultry.
  • Newcastle Disease is regulated by law in many countries and, worldwide, and Newcastle disease is seen as the most important of all diseases that put poultry health at risk.
  • Specific immunoprophylaxis plays the most important role in the prevention of disease and vaccines may be either live attenuated strains administered as aqueous suspensions of previously freeze-dried preparations (with the addition of adequate protective substances), or inactivated preparations, usually in the form of oil emulsions.
  • IBD Infectious bursal disease
  • Gumboro disease is an acute and highly contagious viral disease of young chickens, characterised by significant damages of the bursa of Fabricius and immunosuppression.
  • a two-segment, two- chain KNA virus of the family Birnaviridae causes the disease.
  • Control and prevention of IBD is possible only by timely vaccination and implementation of bio- protection measures.
  • the use of mild, live vaccines prevailed till the end of 1980s when unsuccessful results of vaccination were reported worldwide.
  • Vaccines containing vaccinal strains of intermediary virulence were subsequently introduced (Lukert D. P., Y. M. Saif (2003): Infectious Bursal Disease. Pages 161-179. In: Diseases of Poultry. 11th ed., Editor-in-Chief Saif Y. M. Blackwell Publishing, Ames, Iowa, USA).
  • Animal and avian viruses are highly sensitive and therefore the virus is generally combined with various substances that provide a protection of the virus during freeze drying; this provides a final product that is in the form of a cake containing 1-3% water.
  • the manufacturing process may start with a surplus of virus particles.
  • the product can be stabilised with appropriate protective substances. Problems related to the stabilisation of live vaccines for veterinary medicine have been comprehensively described in WO 2007/919128.
  • the manufacturing process for such two-component vaccines consists essentially of the production of two separate viral suspensions which are mixed and then freeze dried.
  • EP 0332241 relates to combined poultry vaccines of which one component is strain Ma5 of the avian infectious bronchitis virus. There are no details in the document about how the vaccines are prepared but it appears to indicate that the viruses are cultivated separately and mixed before administration to the birds.
  • Combined vaccines are advantageous because it is more cost effective to give a single dose of a combined vaccine instead of separate doses of two or more vaccines.
  • the stress associated with vaccination is reduced by having a single dose rather than separate doses. It would be an advantage to provide a combined vaccine in which the viral strains are produced and freeze dried simultaneously as this would significantly reduce manufacturing costs.
  • a process for the preparation of a viral vaccine comprising culturing at least two different viruses in an embryonated avian egg.
  • the egg in which the viruses are cultured will be a chicken egg, especially an SPF chicken egg.
  • the egg may be inoculated with a mixture of the at least two different viruses.
  • the process may comprise inoculating the egg with a first virus and subsequently with additional viruses. Inoculation may take place about 8 to 14 days after fertilization, typically about 10 to 11 days after fertilization.
  • one of the at least two different viruses is Newcastle disease virus (NDV) or infectious bursal disease virus (IBDV) and in a particularly preferred embodiment of the invention, both NDV and IBDV are cultured in the embryonated egg.
  • NDV Newcastle disease virus
  • IBDV infectious bursal disease virus
  • suitable strains of NDV for use in vaccines are lentogenic strains, particularly the La Sota and Bl strains.
  • Strain VMG 91 is a particularly suitable IBDV strain.
  • the eggs are inoculated with the virus in the form of suspensions of suitable dilution.
  • the NDV suspension may have a titre range of from 10° to 10 6 EID 50 / 0.1ml, preferably from 10 3 5 to IO 45 EID 50 / 0.1ml.
  • the IBDV suspension may contain virus with TCID5 0 in the range of 10° to 10 6 per 0.1ml, preferably from 10 3 5 to 10 45 TCID 50 / O.lmlN
  • the egg is preferably incubated for at least 72 hours. During this time, it is preferred that candling of the egg is carried out at 24 hour intervals to determine viability.
  • the viral products are harvested to obtain a viral suspension comprising the at least two different viruses.
  • the different viruses may replicate in different parts of the egg.
  • NDV is one of the viruses
  • replication occurs mainly in allantoic fluid and, to a lesser extent, in embryos.
  • IBDV occurs mainly in embryos and to a lesser extent in the allantoic fluid.
  • both the embryos and the allantoic fluid are harvested. Generally, the allantoic fluid and the embryos will be collected separately.
  • Embryos may be collected and homogenised with the addition of a buffer solution, especially a phosphate buffer to form a homogenised embryo suspension (one part of embryos and two parts of buffer solution).
  • the homogenised embryo suspension may be stored at a temperature of 2-8°C until needed.
  • the suspension has a virus titre of at least 10 6 ' 5 TCIDso/ml but more preferably at least 10 795 TCID 50 /ml.
  • Allantoic fluid may be collected according to known methods.
  • the virus cultivated in the allantoic fluid is Newcastle Disease virus
  • the allantoic fluid has a virus titre of at least IO 85 EIDso/ml but more preferably at least 10 9 - 83 EID 50 AnI. and may be mixed with a stabiliser solution after collection to form a stabilised allantoic fluid solution.
  • a particularly preferred stabiliser solution comprises an aqueous solution of polyvinylpyrrolidone, sodium glutamate and Bacto peptone.
  • the stabiliser solution also comprises potassium dihydrogen phosphate and potassium hydroxide.
  • This stabilising solution forms a further aspect of the invention.
  • the stabiliser solution may comprise from 0.5 to 10% w/v polyvinylpyrrolidone, 1 to 30% w/v bactopeptone and 0.5 to 10% w/v sodium glutamate in water, preferably purified water
  • the stabiliser solution comprises an aqueous solution containing about 2% w/v polyvinylpyrrolidone, about 10% w/v bactopeptone and about 2% w/v sodium glutamate.
  • the stabiliser solution may also contain additional ingredients, for example potassium dihydrogen phosphate, which may be present in an amount of from about 0.5 to 10% w/v, more usually about 0.5 to 3% w/v and typically about 1.4% w/v; and potassium hydroxide, which may be present in an amount of from about 0.1 to 5% w/v, more preferably about 0.1 to 1% w/v and typically about 0.4% w/v.
  • potassium dihydrogen phosphate which may be present in an amount of from about 0.5 to 10% w/v, more usually about 0.5 to 3% w/v and typically about 1.4% w/v
  • potassium hydroxide which may be present in an amount of from about 0.1 to 5% w/v, more preferably about 0.1 to 1% w/v and typically about 0.4% w/v.
  • the stabilised allantoic fluid solution may comprise allantoic fluid and stabiliser in a volume ratio of from 4:1 to 1:4 allantoic fluid to stabiliser, preferably 4:1 to 1:1 allantoic fluid to stabiliser and usually about 2.5:1 allantoic fluid to stabiliser.
  • the mixture of harvested allantoic fluid and stabiliser solution may be frozen and stored until needed.
  • the stabilised allantoic fluid solution may be mixed with homogenised embryo suspension to form a freeze drying mixture and therefore, in a further aspect of the invention, there is provided a freeze drying mixture comprising 15-45% v/v allantoic fluid, 15-45% v/v homogenised embryo suspension and 5-30% v/v stabiliser solution, with the balance being water.
  • the homogenised embryo suspension is as described above and comprises a suspension of homogenised embryos in a phosphate buffer such as phosphate buffered saline.
  • the stabiliser solution is also as defined above.
  • a preferred freeze drying mixture comprises about 30% v/v allantoic fluid, 30% v/v homogenised embryo suspension, 20% v/v stabiliser solution and 20% v/v water.
  • the freeze drying mixture may be prepared by mixing stabilised allantoic fluid solution as described above with homogenised embryo suspension; and adding purified sterile water. Optionally, further stabilising solution may also be added to the mixture.
  • a typical freeze drying mixture may comprise 42% v/v stabilised allantoic fluid solution (containing 12% v/v stabiliser solution), 30% v/v homogenised embryo suspension; 8% stabiliser solution; and 20% water.
  • Each litre of freeze drying mixture is sufficient to provide approximately 10 6 doses of vaccine.
  • freeze dried vaccine After freeze drying, the freeze dried vaccine must be reconstituted with water before use.
  • the amount of water added to the freeze dried vaccine will depend upon the intended method of administration and the age of the birds to be treated.
  • the water is preferably sterile.
  • each thousand doses will typically be added to 5-25 litres of water.
  • each 1000 doses is added to 10 1 of water and when the vaccine is intended for 2-3 week old birds, each 1000 doses is added to 20 1 of water.
  • each dose may be added to ImI of water (2500 doses in 2.5 1 of water).
  • ImI of water 2500 doses in 2.5 1 of water.
  • a vaccine composition comprising IBDV in a titre of at least 10 20 TCID 50 per dose and NDV in a titre of at least 10 5 0 EID50 per dose.
  • the vaccine composition comprises IBDV in a titre of at least 10 3 0 TCID 50 per dose and NDV in a titre of at least 10 60 EID 50 per dose.
  • a typical dose comprises the following constituents
  • lentogenic strains of NDV are preferred, particularly the La Sota and Bl strains.
  • Strain VMG 91 is a particularly suitable IBDV strain.
  • Figure 1 is a plot showing the HI-titre of specific anti-NDV antibodies in the blood serum of chickens after vaccination with either two-component vaccine against IBD and ND or with ND vaccine.
  • Figure 2 is a plot of ELISA-titre of specific anti-IBDV antibodies in the blood serum of chickens after vaccination with either two-component vaccine against IBD and NV or with vaccine against IBD.
  • Embryonated SPF eggs (VaIo Lohmann) were incubated for 11 days at 37.5°C and 55% humidity with turning of eggs 5 times a day.
  • Newcastle disease virus (NDV) and Gumboro disease virus (GDV) were propagated in embryonated chicken eggs for 72 hours. Candling of eggs was done every 24 hours.
  • Titre of ND virus was 10 4 ' 3 EID 50 /0,l ml
  • Titre of IBD virus was 10 3 9 TCID 50 /0,l ml 3.
  • a mixture of NDV and IBDV was prepared - 5 ml of each inoculum of virus 1 and 2 were mixed.
  • Groups of embryonated SPF eggs were inoculated as follows: Group 1 - Each of 40 embryonated SPF eggs was inoculated with 0.1 ml of mixture of NDV and IBDV.
  • Group 3 Each of 10 embryonated SPF eggs was inoculated with 0.1 ml of ND virus Groups 2 and 3 were inoculated with individual viruses for the control of virus growth.
  • Allantoic fluid was taken separately from each egg containing a live embryo or an embryo that had died between 24 and 72 hours after inoculation, and a haemagglutination (HA) titre of NDV was determined in each.
  • HA haemagglutination
  • Embryos were sorted according to group and according to whether they were alive or dead after 72 hours. Embryos from groups 1, 2, and 3 which were live at 72 hours were designated as groups 1/1, 2/1 and 3/1 while embryos which were dead after 72 hours were designated as groups 1/2, 2/2 and 3/2. After the harvest, the embryos were homogenised with the addition of phosphate buffer solution.
  • 25 ⁇ l of PBS was added to all wells of a microtiter plate in all wells.
  • 25 ⁇ l of virus suspension was added and mixed then 25 ⁇ l of the mixture was transferred to the second row and mixed, following which 25 ⁇ l was added to the next row.
  • the process was continued until the 12 th row obtaining dilutions of 1:2; 1:4; 1:8; 1:16; 1:2048; 1:4096, i.e. of between 1.-2 1 and 1:2 12 .
  • TCID 5 0 is defined as the tissue culture infectious dose which will infect 50% of the challenged cells.
  • a primary culture of chicken fibroblasts was grown on 48-well microtitre plates.
  • the plates were allowed to stand at room temperature for 30 minutes to allow adsorption to take place, after which 0.5ml of modified Eagles medium (MEM) was added to each well.
  • MEM modified Eagles medium
  • the wells were then covered and allowed to stand at a temperature of 37 0 C.
  • the results were read under a microscope after 24 hours and every subsequent 24 hours until a final reading was taken after 144 hours.
  • Table 1 show that the virus suspension obtained from Group 1/1 had a similar HA titre to the virus suspension obtained from Group 3/1 and a similar TCID50 to the virus suspension obtained from Group 2/1.
  • a stabiliser solution was prepared as follows:
  • a mixture for freeze drying was prepared as follows
  • Filling volume for a vial of 2500 doses was 2.5 ml of freeze-drying mixture per vial. Edwards freeze-drier was used.
  • NDV -virus titre >10 60 EID 50 /dose
  • the vaccine was stored in a cooling chamber at 2-8 0 C.
  • Example 3 Accelerated Stability Test Samples of the vaccine were stored in a thermostatic chamber at 37°C.
  • Samples of the vaccine were collected after 24, 72 and 168 hours and then kept in a cooling chamber at 2-8°C until analysis.
  • Example 4 Use of Two-Component Vaccine A study was carried out for the purpose of proving the efficacy of vaccination of chickens with two-component vaccine against infectious bursal disease (Gumboro) and Newcastle disease.
  • the vaccine used in the study was prepared from the strain VMG 91 (intermediary strain) of infectious bursal disease virus (IBDV) and the strain La Sota of Newcastle disease virus (NDV), both propagated in the same chicken egg.
  • IBDV infectious bursal disease virus
  • NDV Newcastle disease virus
  • Hl-titre in the vaccinated chickens increased during the study period, and on day 21 following vaccination it amounted to 2 5 5 in the group vaccinated with two- component vaccine, 2 469 respectively, in the group vaccinated with Newcastle disease vaccine.
  • Chickens of the control group and Group 3 (vaccinated with IBD vaccine) were free from specific anti-NDV antibodies.
  • ELISA-titre in the vaccinated chickens increased during the study period and also in the group vaccinated with two-component vaccine and on day 28 post vaccination it amounted to 2785, 2995 respectively, in the group vaccinated with infectious bursal disease vaccine.
  • Chickens of the control group and Group 1 were free from specific anti-IBDV antibodies.
  • HI-titre of anti-NDV antibodies was found to be higher in chickens vaccinated with two-component vaccine against infectious bursal disease and Newcastle disease in comparison with that recorded in chickens vaccinated only against Newcastle disease. However, there were no statistically significant differences between titre values.
  • ELISA-titre of anti-IBDV antibodies was insignificantly higher in the group of chickens vaccinated with a single component vaccine against IBD in comparison with that recorded in chickens vaccinated with two-component vaccine against infectious bursal disease and Newcastle disease.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Virology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
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  • General Engineering & Computer Science (AREA)
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Abstract

L'invention porte sur un procédé de préparation d'un vaccin viral à plusieurs composants qui comporte la culture d'au moins deux espèces virales différentes dans un embryophore.
PCT/HR2008/000024 2007-07-25 2008-07-08 Procédé de préparation d'un vaccin viral Ceased WO2009013551A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0714489A GB2451428A (en) 2007-07-25 2007-07-25 Process for preparing an in ovo combination viral vaccine
GB0714489.2 2007-07-25

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WO2009013551A2 true WO2009013551A2 (fr) 2009-01-29
WO2009013551A3 WO2009013551A3 (fr) 2009-03-19

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012129872A1 (fr) * 2011-03-25 2012-10-04 天津济命生生物科技有限公司 Produit biologique et son procédé de préparation
WO2012129873A3 (fr) * 2011-03-25 2012-11-22 天津济命生生物科技有限公司 Procédé pour la préparation d'un produit biologique antitumoral

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5037650A (en) * 1988-03-08 1991-08-06 Akzo N.V. Live combination vaccine
IL101183A (en) * 1992-03-09 1996-01-19 Abic Ltd Color-coded inactivated poultry vaccines
RU2058155C1 (ru) * 1993-02-09 1996-04-20 Всероссийский научно-исследовательский институт ветеринарной вирусологии и микробиологии Способ получения ассоциированной вирусвакцины против инфекционного ларинготрахеита и ньюкаслской болезни птиц
US6048535A (en) * 1997-06-12 2000-04-11 Regents Of The University Of Minnesota Multivalent in ovo avian vaccine
JP2002532559A (ja) * 1998-12-21 2002-10-02 アクゾ・ノベル・エヌ・ベー 不活化ｗ／ｏ乳濁液アジュバント化ワクチンの産生法
US6592869B2 (en) * 1999-08-24 2003-07-15 Teva Pharmaceutical Industries, Ltd. Vaccine composition and method of using the same
ZA200104596B (en) * 2000-06-09 2001-12-06 Akzo Nobel Nv IBDV strain for in ovo administration.
UA16452U (en) * 2006-01-30 2006-08-15 Instex And Clinical Veterinary Method for producing live lyophilized associated vaccine against infectious bursal disease (gamboro disease) and newcastle disease

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012129872A1 (fr) * 2011-03-25 2012-10-04 天津济命生生物科技有限公司 Produit biologique et son procédé de préparation
WO2012129873A3 (fr) * 2011-03-25 2012-11-22 天津济命生生物科技有限公司 Procédé pour la préparation d'un produit biologique antitumoral

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GB0714489D0 (en) 2007-09-05
WO2009013551A3 (fr) 2009-03-19

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