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RU2012117014A - METHOD FOR PRODUCING RECOMBINANT CAPSIDE PROTEIN OF HEPATITIS VIRUS E. RECOMBINANT VACCINE FOR PREVENTION OF VIRAL HEPATITIS E. - Google Patents

METHOD FOR PRODUCING RECOMBINANT CAPSIDE PROTEIN OF HEPATITIS VIRUS E. RECOMBINANT VACCINE FOR PREVENTION OF VIRAL HEPATITIS E. Download PDF

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Publication number
RU2012117014A
RU2012117014A RU2012117014/10A RU2012117014A RU2012117014A RU 2012117014 A RU2012117014 A RU 2012117014A RU 2012117014/10 A RU2012117014/10 A RU 2012117014/10A RU 2012117014 A RU2012117014 A RU 2012117014A RU 2012117014 A RU2012117014 A RU 2012117014A
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recombinant
protein
hepatitis
orf2
rthev
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RU2012117014/10A
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Russian (ru)
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RU2501809C1 (en
Inventor
Михаил Александрович Крымский
Иван Андреевич Борисов
Михаил Симеонович Яковлев
Татьяна Анатольевна Семененко
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Закрытое акционерное общество научно-производственная компания "Комбиотех"
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Priority to RU2012117014/10A priority Critical patent/RU2501809C1/en
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  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

1. Способ получения рекомбинантного капсидного белка вируса гепатита Е генотипа 3 (rtHEV-ORF2), включающий культивирование штамма-продуцента, содержащего последовательность ДНК, кодирующую белок ORF2, в подходящих для экспрессии указанного белка условиях, выделение и очистку целевого белка, отличающийся тем, что культивируют рекомбинантный штамм дрожжей Hansenula polymorpha КБТ-11/pHEV-001, содержащий интегрированную в геном последовательность ДНК, включающую рекомбинантный ген, кодирующий полипептид rtHEV-ORF2 с аминокислотной последовательностью SEQ ID NO:1, под контролем промотора гена МОХ Hansenula polymorpha и ген LEU2 S.cerevisiae в качестве селективного маркера, а после выделения и очистки получают белок rtHEV-ORF2 в виде VLP, состоящей из мономеров с молекулярной массой мономера 56 кДа.2. Рекомбинантная вакцина для профилактики вирусного гепатита Е, содержащая эффективное количество капсидного белка вируса гепатита Е, адъювант и физиологически приемлемый разбавитель, отличающаяся тем, что в качестве антигена вируса гепатита Е вакцина содержит белок rtHEV-ORF2, полученный по способу п.1.3. Рекомбинантная вакцина по п.2, отличающаяся тем, что дополнительно содержит мертиолят.1. A method of producing a recombinant hepatitis E virus capsid protein of genotype 3 (rtHEV-ORF2), comprising cultivating a producer strain containing a DNA sequence encoding an ORF2 protein under conditions suitable for expression of said protein, isolating and purifying the target protein, characterized in that cultivate a recombinant strain of Hansenula polymorpha KBT-11 / pHEV-001 containing the DNA sequence integrated into the genome, including the recombinant gene encoding the rtHEV-ORF2 polypeptide with the amino acid sequence of SEQ ID NO: 1, under warning light gene promoter and MOX Hansenula polymorpha LEU2 S.cerevisiae gene as a selectable marker, and after isolation and purification rtHEV-ORF2 protein in a VLP, consisting of monomers with a molecular mass of 56 kDa.2 monomer. Recombinant vaccine for the prevention of viral hepatitis E, containing an effective amount of the capsid protein of the hepatitis E virus, an adjuvant and a physiologically acceptable diluent, characterized in that the vaccine contains the rtHEV-ORF2 protein obtained by the method of clause 1.3 as an antigen of the hepatitis E virus. The recombinant vaccine according to claim 2, characterized in that it further comprises thiolate.

Claims (3)

1. Способ получения рекомбинантного капсидного белка вируса гепатита Е генотипа 3 (rtHEV-ORF2), включающий культивирование штамма-продуцента, содержащего последовательность ДНК, кодирующую белок ORF2, в подходящих для экспрессии указанного белка условиях, выделение и очистку целевого белка, отличающийся тем, что культивируют рекомбинантный штамм дрожжей Hansenula polymorpha КБТ-11/pHEV-001, содержащий интегрированную в геном последовательность ДНК, включающую рекомбинантный ген, кодирующий полипептид rtHEV-ORF2 с аминокислотной последовательностью SEQ ID NO:1, под контролем промотора гена МОХ Hansenula polymorpha и ген LEU2 S.cerevisiae в качестве селективного маркера, а после выделения и очистки получают белок rtHEV-ORF2 в виде VLP, состоящей из мономеров с молекулярной массой мономера 56 кДа.1. A method of producing a recombinant hepatitis E virus capsid protein of genotype 3 (rtHEV-ORF2), comprising cultivating a producer strain containing a DNA sequence encoding an ORF2 protein under conditions suitable for expression of said protein, isolating and purifying the target protein, characterized in that cultivate a recombinant strain of Hansenula polymorpha KBT-11 / pHEV-001 containing the DNA sequence integrated into the genome, including the recombinant gene encoding the rtHEV-ORF2 polypeptide with the amino acid sequence of SEQ ID NO: 1, under warning light gene promoter and MOX Hansenula polymorpha LEU2 S.cerevisiae gene as a selectable marker, and after isolation and purification rtHEV-ORF2 protein in a VLP, consisting of monomers with a molecular mass of 56 kDa monomer. 2. Рекомбинантная вакцина для профилактики вирусного гепатита Е, содержащая эффективное количество капсидного белка вируса гепатита Е, адъювант и физиологически приемлемый разбавитель, отличающаяся тем, что в качестве антигена вируса гепатита Е вакцина содержит белок rtHEV-ORF2, полученный по способу п.1.2. Recombinant vaccine for the prevention of viral hepatitis E, containing an effective amount of a capsid protein of the hepatitis E virus, an adjuvant and a physiologically acceptable diluent, characterized in that the vaccine contains the rtHEV-ORF2 protein obtained by the method of claim 1 as an antigen of the hepatitis E virus. 3. Рекомбинантная вакцина по п.2, отличающаяся тем, что дополнительно содержит мертиолят. 3. The recombinant vaccine according to claim 2, characterized in that it further comprises thiolate.
RU2012117014/10A 2012-04-27 2012-04-27 Method for obtaining recombinant core protein of hepatitis e virus and recombinant vaccine for prophylaxis of hepatitis e virus RU2501809C1 (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023231149A1 (en) * 2022-05-25 2023-12-07 Xuzhou Medical University Recombinant orthohepevirus genotype c1 orf2 protein and preparation method therefor and use thereof

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RU2745558C2 (en) * 2016-12-27 2021-03-29 Федеральное государственное бюджетное научное учреждение "Федеральный научный центр исследований и разработки иммунобиологических препаратов им. М.П. Чумакова РАН" Method of producing recombinant capsid protein of hepatitis e virus

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US6291641B1 (en) * 1988-06-17 2001-09-18 Genelabs Technologies, Inc. Hepatitis E virus antigens and uses therefor
US6455492B1 (en) * 1988-06-17 2002-09-24 Genelabs Technologies, Inc. Hepatitis E virus vaccine and method
US6458562B1 (en) * 1998-04-09 2002-10-01 The United States Of America As Represented By The Secretary Of Health And Human Services Recombinant proteins of a Pakistani strain of hepatitis E and their use in diagnostic methods and vaccines
RU2252748C1 (en) * 2004-02-11 2005-05-27 Солонский Александр Владимирович Method for treating viral hepatitis
CN101062941A (en) * 2007-04-27 2007-10-31 东南大学 Recombined hepatitis E hepatitis virus protein, preparation method and usage thereof

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023231149A1 (en) * 2022-05-25 2023-12-07 Xuzhou Medical University Recombinant orthohepevirus genotype c1 orf2 protein and preparation method therefor and use thereof

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