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Inactivated vaccine for preventing monkey from SV40 virus infection and preparation method

An inactivated vaccine, virus infection technology, applied in the direction of antiviral agents, virus antigen components, etc.

Inactive Publication Date: 2008-01-09
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But at present, there is no vaccine against monkey SV40 infection and its immunological reports at home and abroad

Method used

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0017] 1. Purification

[0018] (1) SV40 virus proliferates in Vero cells. After the Vero cells form a monolayer, inoculate SV40 virus at 3m.o.i, absorb and suspend culture for 3-5 days. Freeze and thaw 4 to 5 times, shake vigorously several times during this period; then combine SV40 of the same generation.

[0019] (2) After merging the virus, centrifuge at 3000rpm, collect the supernatant for 10min, slowly add NaCl, PEG6000 to make the final concentration 0.5Mg / (WN), collect the precipitate at room temperature for 1h, 4°C for 36h, 10000rpm for 25min, dissolve in 25mMTris, 137mMNaCl, 5mMKCl, l0.7mMNa 2 PO 4 , 25mMMgCl 2 , In the TD solution formed. In an ice bath, slowly add sodium deoxycholate and Triton X-100 to make the final concentrations of the cell solution reach 0.5% and 1.0%, respectively.

[0020] (3) Sucrose was dissolved in pH7.4, 0.02mol / L Tris solution, the density of the upper layer was 20% (W / W), and the density of the lower layer was 75% (W / W); then dis...

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Abstract

The invention is concerned with a kind of inactivated vaccine and preparation method to anti-monkey SV40 virus, belonging to animal vaccine. It carries multiplication, inactivation and purification to prepare inactivated vaccine of anti-monkey SV40 virus. This vaccine is SV40 virus and adjuvant inactivated with beta-Propiolactone and the cubage of them is 2:1 to 3:2. Mix the said component with buffer solution and get Tween-20 with 0.03 to 0.05 mg / ml and glycine with 2.0 to 4.0mg / ml, and the virus content is 104.6 to 5.2TCID50 / ml. Purificate the SV40 virus liquid with 0 percent CPE to collect virus, inactivate and stop SV40 virus with beta-Propiolactone and use adjuvant absorb inactivated SV40virus. The specific antibody caused of small rat can identify SV40virus with security. The tested monkey can produce neutralizing antibody with singularity, and the neutralizing antibody with singularity can stop the infection of SV40virus to cell culturing matter during the test. This inactivated vaccine maybe has natural anti-infection function.

Description

Technical field [0001] The invention belongs to animal vaccines. Background technique [0002] SV40 virus belongs to simian vacuolar virus 40, which infects rhesus macaques in a natural state. Infection with SV40 virus can cause fatal related diseases including pneumonia and kidney disease, which is a very great potential threat to both humans and monkeys. In this regard, overseas studies have focused on the tumorigenic mechanism of SV40 and anti-tumor immunity. By expressing SV40T antigen and constructing SV40T DNA vaccine as a tumor vaccine, immunosuppression of tumors is carried out, especially by removing the binding site of T gene Rb and P53. The point is to improve the safety of the vaccine, and the vaccine has entered clinical research. Domestic research focuses on the construction of eukaryotic expression vectors using the SV40 viral promoter. There are few domestic and foreign reports on the research on SV40 inactivated vaccine for monkeys and other forms of vacci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61P31/12
Inventor 孙茂盛李鸿钧和占龙张光明谢天宏曹增孙雯佳孙强明唐永明蒋鸿超
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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