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Method for preparing baculovirus-loaded cyprinid herpesvirus type II DNA vaccine by freeze drying

A DNA vaccine, baculovirus technology, applied in DNA/RNA vaccination, intact cell/virus/DNA/RNA components, double-stranded DNA virus, etc., to prevent impact, improve application effect, and good stability

Pending Publication Date: 2021-11-23
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previously, the inventor used -80°C pre-freezing overnight, and then carried out low-pressure step-by-step drying, and the effect of the obtained freeze-dried powder was slightly improved. However, after several repeated tests, it was found that the stability of the product performance of this process needs to be improved.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Example 1 Construction method of cyprin herpesvirus type II DNA vaccine based on baculovirus vector and its injection immune effect

[0013] Refer to Example 3 of CN2018105344579, the first 6 steps are consistent with it, and step (7) is as follows:

[0014] (7) Take the P2 generation recombinant virus with insect needles, puncture and inoculate the initial pupation from the intersegmental membrane, protect at 25°C for 5 days, collect the recombinant virus to infect silkworm pupae, add 0.01mol / L at a ratio of 1:10 (W / V) Phosphate buffered saline, homogenate. After the homogenate is filtered through 8 layers of gauze, the filtrate is obtained; the above is also the content disclosed in the third embodiment of CN2018105344579; the inventive steps of this embodiment are as follows: the filtrate is freeze-dried to prepare silkworm chrysalis freeze-dried powder, and the water content of the freeze-dried powder The yield is less than 5%; the freeze-dried powder is stored at ...

Embodiment 2

[0017] The embodiment was repeated once or twice, and the results showed that the protection rates of oral immunization were 67% and 68%, respectively. Illustrate, the stability of the inventive method is better.

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Abstract

The invention relates to a method for preparing baculovirus-loaded cyprinid herpesvirus type II DNA vaccine by freeze drying. The recombinant virus BmNPV-FA-D4ORF is inoculated to bombyx mori, then the diseased bombyx mori is homogenized and freeze-dried, freeze-dried powder is prepared, and the cyprinid herpesvirus II type DNA vaccine is obtained; and the freeze-drying step comprises the steps of pre-freezing overnight at -80 DEG C, and then carrying out low-pressure stepped heating drying. The freezing method is limited, and the application effect is further improved on the basis that the existing recombinant virus BmNPV-FA-D4ORF is inoculated to the bombyx mori and then the diseased bombyx mori is homogenized.

Description

technical field [0001] The invention relates to the technical field of viral genetic engineering, in particular to a method for preparing baculovirus-carried carp herpesvirus type II DNA vaccine by freeze-drying. Background technique [0002] There are many ways to immunize fish. Among them, the injection immunization effect is good but the workload is heavy, and the oral vaccine damages little damage but is easily affected by digestion. In order to solve this problem, the prior art CN2018105344579 discloses a baculovirus-based carp Herpes virus type II DNA vaccine and its construction method and application, specifically the following steps, (1) clone the promoter into the SmaI / XhoI site of pFSATBacDual to obtain the pFSATBacDual-FA vector; (2) clone the fusion DNA sequence into pFSATBacDual- The XhoI / KpnI site of FA was used to obtain the pFSATBacDual-FA-D4ORF plasmid; (3) Transform the pFSATBacDual-FA-D4ORF plasmid into DH10 / Bac competent cells, spread on LB agar medium p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/245A61K9/19A61P31/22
CPCA61K39/12A61K9/19A61P31/22C12N2710/16034A61K2039/53
Inventor 徐龙行张星胡小龙刘波贡成良李坤
Owner SUZHOU UNIV
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