A kind of protein vaccinia vaccine and its efficacy detection method

A vaccinia vaccine and protein technology, applied in the field of vaccines, can solve problems such as unclear mechanism of action, and achieve the effect of preventing smallpox virus infection and excellent immune effect

Active Publication Date: 2021-10-22
CARBIOGENE THERAPEUTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the ability of many recombinant proteins to activate the host immune response is weak, so protein vaccines need to be used together with adjuvants to enhance the immunogenicity of the antigen
It is generally believed that the type of adjuvant directly determines the effect of immunization and the type of immune response, but the mechanism of action is still unclear

Method used

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  • A kind of protein vaccinia vaccine and its efficacy detection method
  • A kind of protein vaccinia vaccine and its efficacy detection method

Examples

Experimental program
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Effect test

preparation example Construction

[0055] The preparation method of above-mentioned protein vaccinia vaccine comprises the following steps:

[0056] 1) Spin column treatment: regenerate and balance the resin in the spin column after the first use and after each use, the specific method is:

[0057] 1.1) Return the spin column to room temperature;

[0058] 1.2) Unscrew the switch at the bottom of the spin column, unscrew the top cap, place the spin column in the collection tube, centrifuge, remove the storage liquid in the spin column, and discard the liquid in the collection tube;

[0059] 1.3) Remove the top cap, plug the bottom plug, add 0.15-0.25 N NaOH to the spin column for regeneration, put on a new top cap, invert the spin column and mix several times until the resin in the spin column is completely suspended in the liquid , standing overnight at room temperature;

[0060] 1.4) Loosen the top cap, remove the bottom plug, place the spin column in the collection tube, centrifuge, and discard the liquid i...

Embodiment 1

[0085] 1. The endotoxin removal steps of virus recombinant protein:

[0086] Use the "Pierce™ High Capacity Endotoxin RemovalSpin Columns, 0.25 mL" kit from Thermo Scientific for endotoxin removal. The following is the specific operation procedure:

[0087] 1. The resin in the spin column needs to be regenerated and balanced after the first use and after each use. The specific operation is as follows:

[0088] 1.1. Return the spin column to room temperature.

[0089] 1.2. Unscrew the switch at the bottom of the spin column, unscrew the top cap, place the spin column in the collection tube, and centrifuge at 500xg for 1 min to remove the storage solution in the spin column. Discard the liquid in the collection tube.

[0090] 1.3. Take off the top cap, plug the bottom plug, add 0.2 N NaOH to the spin column for regeneration, put on a new top cap, turn the spin column upside down and mix several times until the resin in the spin column is completely suspended in the liquid . ...

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Abstract

The invention relates to the field of vaccines, and discloses a protein vaccinia vaccine and a method for detecting its effectiveness. The protein vaccine contains viral recombinant protein D8L expressed by yeast and with a purity >90% of endotoxin removed. The preparation method is as follows: expressing yeast with a purity >90% 90% of the recombinant protein D8L of vaccinia virus has been treated to remove endotoxin to obtain a recombinant protein D8L solution with an endotoxin content <5 EU / ml. Efficacy testing method is as follows: the recombinant protein D8L solution that removes endotoxin is injected subcutaneously twice to immunize mice. After 21 days, specific antibodies and neutralizing antibodies can be detected in the mouse serum, indicating that the mice have acquired anti-vaccinia virus immunity. immune protection. The protein vaccine of the present invention does not require additional vaccine adjuvants, and can theoretically replace traditional live vaccinia virus vaccines, and play a role in resisting pox virus infections such as smallpox virus.

Description

technical field [0001] The invention relates to the field of vaccines, in particular to a protein vaccinia vaccine and a method for detecting its efficacy. Background technique [0002] Vaccinia virus is a member of the poxvirus family Orthopoxvirus, which includes smallpox virus, monkeypox virus, vaccinia virus and mousepox virus. Vaccinia virus has a complex and large double-stranded DNA genome of up to 200Kb, which can encode most of the genes required for viral cytoplasmic replication. Vaccinia virus is the most studied of the poxvirus family, and live vaccinia virus vaccines have been used to immunize against smallpox virus since the late 1970s. Today, although smallpox has been extinct, the decline in the vaccination rate of vaccinia has given terrorists the opportunity to recreate the smallpox virus, endangering public safety. Widespread vaccination is considered an effective means of protection against this threat. However, current vaccines based on live vaccinia ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/285A61P31/20G01N33/68
CPCA61K39/12A61K2039/552A61P31/20C12N2710/24134G01N33/6854
Inventor 杨文君其他发明人请求不公开姓名
Owner CARBIOGENE THERAPEUTICS CO LTD
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