Recombinant adenovirus and tetravalent adenovirus vaccine and preparation method thereof

A technology of recombinant adenovirus and adenovirus, which is applied in the field of genetic engineering, can solve the problem of no prevention, and achieve the effect of similar genome copy number and replication efficiency

Active Publication Date: 2017-01-11
GUANGZHOU GIR MEDICINE CO LTD +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this vaccine can only prevent human type 4 and type 7 adenovirus infection, an

Method used

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  • Recombinant adenovirus and tetravalent adenovirus vaccine and preparation method thereof
  • Recombinant adenovirus and tetravalent adenovirus vaccine and preparation method thereof
  • Recombinant adenovirus and tetravalent adenovirus vaccine and preparation method thereof

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Embodiment 1 6

[0034] Example 1 Preparation of hexon replacement type recombinant adenovirus rAd3H14, rAd3H55 and rAd3H7

[0035] The backbone plasmid pBRAd△E3GFP obtained by the inventor in the early stage (published documents are Zhang Q, Su X, Seto D, Zheng BJ, Tian X, Sheng H, Li H, Wang Y, Zhou R. Construction and characterization of a replication-competent human adenovirus type 3-based vector as a live-vaccine candidate and a viral delivery vector. Vaccine 2009; 27(8):1145-53) as the basis, using the full-length hexon gene of Ad7, Ad14 or Ad55 (amplified from The virus strain of Ad7, Ad14 or Ad55, without mutation in the amino acid sequence) replaces the hexon gene of the recombinant type 3 adenovirus to obtain the hexon replacement type recombinant adenovirus rAd3H7, rAd3H14, rAd3H55. The basic construction procedure is as follows: amplify the hexon gene of Ad7, Ad14 or Ad55 by PCR, clone it into the hexon shuttle plasmid pBRHexonL / R, use pBRAd△E3GFP as the backbone plasmid, and use t...

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Abstract

The invention discloses a recombinant adenovirus and tetravalent adenovirus vaccine and a preparation method thereof. The tetravalent recombinant adenovirus vaccine contains a recombinant type 3 adenovirus strain, a recombinant type 7 adenovirus strain, a recombinant type 14 adenovirus strain and a recombinant type 55 adenovirus strain. The preparation method disclosed by the invention comprises the following steps: preparing recombinant shuttle plasmids containing hexon gene segments, and performing in-bacteria homologous recombination with a recombinant human type 3 adenovirus strain, thereby obtaining a recombinant adenoviral genome in which the hexon gene segments are replaced by type 7, type 14 and type 55; transfecting cells, rescuing to obtain recombinant human type 3, 7, 14 and 55 recombinant adenoviruses with different main capsid protein-hexon proteins; purifying, mixing according to the same protein content, and inactivating by using beta-propiolactone, thereby obtaining the tetravalent adenovirus vaccine. The tetravalent adenovirus vaccine is capable of inducing neutralizing antibody responses to four types of serotype adenoviruses, and the neutralizing titer is 500-1000.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and specifically relates to a recombinant adenovirus, a quadrivalent human adenovirus vaccine using human type 3 adenovirus (HAdv3) as a carrier and a preparation method thereof. Background technique [0002] Human Adenovirus (HAdV) is a type of pathogen first isolated and identified from patients with acute respiratory diseases in 1953. It can infect human respiratory tract, gastrointestinal tract, urinary system or eyes and other tissues, leading to acute respiratory infection. (ARI), acute gastroenteritis, nephritis, keratoconjunctivitis and many other diseases. Adenoviruses are widespread and spread rapidly. In recent years, there have been outbreaks and epidemics in many countries and regions, which seriously threaten public health security. Adenovirus-infected patients are mainly infants and infirm people. Adenovirus outbreaks occur from time to time in densely populated kinder...

Claims

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Application Information

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IPC IPC(8): C12N7/01C12N15/861A61K39/295A61K39/235A61P31/20
CPCA61K39/12C07K14/005C12N15/86C12N2710/10021C12N2710/10022C12N2710/10034C12N2710/10043
Inventor 田新贵周荣蒋再学马强李潇周志超
Owner GUANGZHOU GIR MEDICINE CO LTD
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