Varicella-zoster virus r-gE fusion protein, recombinant varicella-zoster vaccine and preparation method and application of varicella-zoster virus r-gE fusion protein and recombinant varicella-zoster vaccine

A herpes zoster virus and fusion protein technology, applied in the field of biomedicine, can solve problems such as weak cellular immunity, achieve simple preparation process, convenient use, prevent or improve the effect of neuralgia

Active Publication Date: 2021-11-23
ANHUI ZHIFEI LONGCOM BIOPHARM CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0017] But this kind combination vaccine has following shortcoming: the patent that application number is 200410033878.1 adopts aluminum hydroxide combined with CpG as the

Method used

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  • Varicella-zoster virus r-gE fusion protein, recombinant varicella-zoster vaccine and preparation method and application of varicella-zoster virus r-gE fusion protein and recombinant varicella-zoster vaccine
  • Varicella-zoster virus r-gE fusion protein, recombinant varicella-zoster vaccine and preparation method and application of varicella-zoster virus r-gE fusion protein and recombinant varicella-zoster vaccine
  • Varicella-zoster virus r-gE fusion protein, recombinant varicella-zoster vaccine and preparation method and application of varicella-zoster virus r-gE fusion protein and recombinant varicella-zoster vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1 r-gE fusion protein eukaryotic expression plasmid construction

[0058] The antigen used in a recombinant varicella-zoster vaccine is in the form of r-gE fusion protein, including VZV gE extracellular sequence and ORF7 protein fragment. VZV gE selects the N-segment extracellular sequence of Dumas strain (GeneBank No.: NP_040190.1), and ORF7 sequence (GeneBank No.: AXF42671.1) selects the N-segment amino acid sequence. The amino acid sequence of the r-gE fusion protein is shown in SEQ ID NO.1; the theoretical molecular weight of the protein is about 66kD (the protein undergoes intracellular modification after expression, and the actual molecular weight may increase).

[0059] The r-gE fusion protein gene was synthesized by Nanjing GenScript Co., Ltd., and the delivered product was a pUC57 plasmid vector. HindIII and EcoR1 restriction sites were introduced at both ends of the expression framework, and the target gene was codon-optimized according to the CHO c...

Embodiment 2

[0061] Embodiment 2 liposome adjuvant compatible varicella-zoster vaccine

[0062] The vaccine is prepared by r-gE fusion protein and liposome containing agonist, and the specific preparation method is as follows.

[0063] 2.1. Preparation of antigenic protein

[0064] Referring to the method recorded in the "Molecular Cloning Experiment Guide", the specific preparation method of the antigen r-gE fusion protein is as follows:

[0065] S1. Transfection of the recombinant plasmid: after linearizing the pEE12.4-r-gE plasmid constructed by the method described in Example 1, transfect it into CHO cells using lip2000 liposomes (refer to the lip2000 liposome transfection instructions); After 48 hours of transfection, take the cell supernatant and use Western Blot to detect the expression of r-gE fusion protein; after 48 hours of transfection, take the cell supernatant and use Western Blot to detect the expression of r-gE fusion protein; , using gE protein mouse monoclonal antibody ...

Embodiment 3

[0082] Embodiment 3 recombinant adenovirus vector varicella-zoster vaccine

[0083] Recombinant adenoviral vector varicella-zoster vaccine (recombinant adenoviral vector vaccine preparation) is recombined from r-gE fusion protein and adenoviral vector, denoted as vaccine 4 (r-gE-Ad), or r-gE fusion The protein is recombined with an adenovirus vector expressing a TLR receptor agonist, and is designated as vaccine 5 (r-gE-R-Ad).

[0084] The specific preparation method is as follows:

[0085] 3.1. Preparation of herpes zoster recombinant adenovirus vector virus

[0086] Experimental materials and sources: The gene sequence of the r-gE fusion protein was synthesized by a commercial company, and the amino acid sequence of the protein is shown in SEQ ID NO.1; DNA Polymerase was purchased from Novozyme, and LR recombinase was purchased from Invitrogen.

[0087] Preparation method: The method described in Example 1 was used to construct the r-gE fusion protein gene expression plasm...

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Abstract

The invention provides a varicella-zoster virus r-gE fusion protein, a recombinant varicella-zoster vaccine and a preparation method and application of the varicella-zoster virus r-gE fusion protein and the recombinant varicella-zoster vaccine. An antigen of the recombinant varicella-zoster vaccine is the varicella-zoster virus r-gE fusion protein, the recombinant varicella-zoster vaccine can be an adjuvant compatible varicella-zoster vaccine which is formed by compatibility of an r-gE fusion protein and an adjuvant, or a recombinant adenovirus vector varicella-zoster vaccine which is formed by gene recombination of the r-gE fusion protein and an adenovirus vector or an adenovirus vector containing a TLR receptor stimulant. The recombinant varicella-zoster vaccine is suitable for preventing or improving varicella-zoster and/or post-varicella-zoster neuralgia; the varicella-zoster virus r-gE fusion protein can further be used for preparing a respiratory syncytial virus and varicella-zoster combined vaccine, and the combined vaccine is formed by compatibility of the r-gE fusion protein, RSV Pre-F protein mixed liquor and the adjuvant; and the prepared combined vaccine can be used for preventing or improving respiratory syncytial virus infection diseases and varicella-zoster.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a varicella-zoster virus r-gE fusion protein, a recombinant varicella-zoster vaccine and a preparation method and application thereof. Background technique [0002] Chickenpox is a highly contagious acute infectious disease caused by the initial infection of varicella-zoster virus. The main route of transmission is respiratory droplets or direct contact, and it mostly occurs in spring and winter. Infants and young children are prone to the disease, and the incidence rate of susceptible children can reach more than 95%, and preschool children aged 2 to 6 are the peak group of the disease. Herpes zoster (HZ) is caused by reactivation of varicella-zoster virus (VZV) during primary varicella infection and is characterized by dermatoid pain and papules; pain usually precedes The rash appears for a few days and can last for months after the rash resolves; the rash tends to appea...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K39/25A61P31/22A61K39/155A61P31/14A61K39/385
CPCC07K14/005C07K14/47A61K39/12A61P31/22A61P31/14A61K39/385C12N2710/16722C12N2710/16734C07K2319/00C12N2760/18534A61K2039/6075A61K2039/70A61K2039/55505A61K2039/55511A61K2039/542A61K2039/543A61K2039/54Y02A50/30
Inventor 谭小东祁碧玉靳正伟庄宗兰潘媛杨文静金香玉程杰彭杰方超杨世龙
Owner ANHUI ZHIFEI LONGCOM BIOPHARM CO LTD
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