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Nanometer antigen particle based on self-assembled ferritin, infectious bursal disease vaccine prepared from nanometer antigen particle, and application

A technology for bursal disease and ferritin, which is applied in the field of infectious bursal disease vaccines, can solve problems such as the recovery of virulence of attenuated strains, and achieve the effects of increasing expression and titer.

Active Publication Date: 2021-03-05
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biggest drawback of using attenuated strain vaccines is the risk of the attenuated strain reverting to virulence in susceptible populations

Method used

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  • Nanometer antigen particle based on self-assembled ferritin, infectious bursal disease vaccine prepared from nanometer antigen particle, and application
  • Nanometer antigen particle based on self-assembled ferritin, infectious bursal disease vaccine prepared from nanometer antigen particle, and application
  • Nanometer antigen particle based on self-assembled ferritin, infectious bursal disease vaccine prepared from nanometer antigen particle, and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Preparation and efficacy testing of IBDV VP2-Ferritin original sequence nanoparticle vaccine

[0070] 1 About the configuration of solution and medium

[0071] For the configuration methods of the solution and medium, refer to relevant reference books (Joseph et al., Molecular Cloning Experiment Guide, Third Edition, 2002; Osper, et al., Molecular Biology Guide, 1998).

[0072] 2 Synthesis of IBDV VP2 structural protein gene sequence and ferritin gene sequence.

[0073] In order to better fuse and express the VP2 structural protein of infectious bursal disease virus and ferritin, the signal peptide analysis software (SignalP) and the transmembrane domain analysis software (TMHMM) were used to analyze the structure of infectious bursal disease virus VP2. According to the amino acid sequence analysis of the protein (GenBank sequence number on NCBI: AVE14454.1), IBDV VP2 structural protein has no signal peptide and transmembrane region, so we choose the complete...

Embodiment 2

[0150] Example 2 The original sequence of IBDV VP2-Ferritin is used for the design of consensus sequence and the preparation and efficacy testing of nanoparticle vaccine after its optimization

[0151] 1 About the configuration of solution and medium

[0152] See Example 1 for the configuration method of the specific solution and culture medium.

[0153] 2 Gene Acquisition of Conserved Sequence of Infectious Bursal Disease Virus VP2 Structural Protein

[0154] The present invention compares the original amino acid sequence of the infectious bursal disease virus VP2 structural protein in Example 1 with other 20 VP2 structural protein amino acid sequences obtained from NCBI to obtain a consensus sequence, which is then optimized. Taking OptimumGene Further TM The technology optimizes the amino acid sequence of IBDV VP2 structural protein, and transforms the optimized amino acid sequence of VP2 structural protein and ferritin monomer subunit amino acid sequence according to the...

Embodiment 3

[0186] Example 3 IBDV VP2-Ferritin-C-O mutant preparation and efficacy testing of nanoparticle vaccine after amino acid single point mutation

[0187] 1 Experimental method

[0188] 1.1 Construction of IBDV VP2-Ferritin-C-O amino acid sequence single point mutant gene

[0189] Based on the results of Example 2, the present invention obtained the IBDV VP2-Ferritin-C mutant, and used the codon-optimized gene sequence of the IBDV VP2-Ferritin-C mutant as a template to design multiple pairs of primers to perform site-directed mutation on the conserved sequence. The site-directed mutagenesis is carried out by fusion PCR method, see Example 1 for the fusion PCR method.

[0190] The mutation sites were IBDV: Y45F, S76T, M85I, L101P, G116R, T128R, S146R, E162D, I184S, Y214H, T228R, Q249H, A275G, T286S, I296N, I308L, G331R, V349E, P375A or R407S. The resulting mutant was named IBDV VP2-Ferritin-C-O-M (Y45F, S76T, M85I, L101P, G116R, T128R, S146R, E162D, I184S, Y214H, T228R, Q249H, A275...

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Abstract

The invention discloses a nanometer antigen particle based on self-assembled ferritin, an infectious bursal disease vaccine prepared from the nanometer antigen particle, and application. Infectious bursal disease virus VP2 structural protein is fused with a self-assembled ferritin nanometer subunit to obtain fusion protein. Parts of the sites of the infectious bursal disease virus VP2 structural protein are mutated, and therefore, the soluble expression quantity and the expression efficiency of a mutant are obviously improved. By use of a prokaryotic expression system and a bombyx mori and AcMNPV-insect cell eukaryotic expression system, recombined protein is expressed, or a recombinant baculovirus is subjected to gene submission in the body of a vertebrate to generate an antigen to induceto generate an antibody. The vaccine provided by the invention shows the infectious bursal disease virus VP2 structural protein on the surface of the cage structure of helicobacter pylori ferritin tocause a universal neutralization antibody capable of resisting the infectious bursal disease, immune potency is improved, an immune range is enlarged, and the vaccine provided by the invention has potential to become a universal vaccine with the cross immune potency.

Description

technical field [0001] The present invention relates to nano-antigen particles based on self-assembled ferritin, in particular to nano-antigen particles fused together by infectious bursal disease virus VP2 structural protein and monomeric ferritin subunits and an infectious method prepared from the nano-particle antigen A bursal disease vaccine belongs to the field of preparation and application of infectious bursal disease vaccines. Background technique [0002] Chicken infectious bursal disease (Infectious bursal disease, IBD, also known as Gamporo disease) is an acute and highly contagious disease that easily infects chickens. The pathogen is infectious bursal disease virus (Infectious bursal disease virus, IBDV), which is highly contagious and has a high incidence rate, almost 100%. IBDV mainly attacks the bursa of Fabricius (Bursa offabricius, BF), the central organ of humoral immunity in chickens, causing bursa offabricius lesions. The bursa of Fabricius is also kno...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/866C12N15/62B82Y40/00A61K39/12A61P31/14
CPCC07K14/195C07K14/415C07K14/435C12N15/86B82Y40/00A61K39/12A61P31/14C12N2720/10034A61K2039/552A61K2039/5258C12N2710/14043C07K2319/00Y02A50/30
Inventor 胡小元李轶女张志芳刘兴健易咏竹张伟业宋浩志
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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