An Inactivated Vaccine Produced Using Attenuated Poliomyelitis Strain

A technology for poliomyelitis and attenuated strains, applied in antiviral agents, viral antigen components, and resistance to vector-borne diseases, etc., can solve the problems of different gene sequences, isoelectric point stability, and low vaccine yields, etc. To achieve the effect of improving immunogenicity, reducing side reactions, and reducing production costs

Active Publication Date: 2019-12-06
SINOVAC BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the attenuated Sabin strain used in sIPV production is essentially different from the wild strain Salk strain used in IPV production, such as the gene sequence, isoelectric point, and stability of the virus. Adaptation to sIPV production results in lower vaccine yields

Method used

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  • An Inactivated Vaccine Produced Using Attenuated Poliomyelitis Strain
  • An Inactivated Vaccine Produced Using Attenuated Poliomyelitis Strain
  • An Inactivated Vaccine Produced Using Attenuated Poliomyelitis Strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The preparation of embodiment 1 cell working seed and the preparation of virus liquid

[0041] 1. Preparation of high-density cell working seeds

[0042] Use 199 culture medium to dilute Vero main cell seed dilution ratio to 60:1, inoculate to 175cm 2 Cells were cultured in square flasks at a temperature of 36.0°C±1°C. After the cells are fully covered with the cell bottle, continue passage for 4 generations according to the ratio of 1:4 to 1:6. The last generation was transferred to five 40-layer cell factories, trypsinized, and centrifuged to remove the supernatant. Use freezing solution containing 10% DMSO to resuspend, and the cell concentration after mixing is 2.5×10 7 pieces / ml. The cell seeds were subjected to gradient cooling in a temperature program cooling device, keeping the temperature down by 1°C per minute. After the temperature drops to -196°C, it is stored in liquid nitrogen, and the working cell seeds are valid for 10 years.

[0043] 2. Preparatio...

Embodiment 2

[0053] Example 2 Purification and Effect Verification of Sabin Strain Poliomyelitis Type I, Type II and Type III Viruses

[0054] 1. The low and high concentrations of the sucrose solution are 25% and 55% respectively. The density gradient centrifugation buffer is 0.5mol / L PBS buffer, and its pH value is 6.5. Use Hitachi CP70MX / ME ultracentrifuge, pump in 100ml ultracentrifugation buffer solution from side hole successively with 100rpm, the sIPV I type (or sIPV II type or sIPV III type) virus ultrafiltration that the embodiment 1 ultrafiltration concentration of about 700ml obtains Liquid, 500ml of low-concentration sucrose solution, and finally pump high-concentration sucrose solution at 100rpm until the liquid flows out of the mesopore, and ultrafiltration and centrifugation at 30,000rpm at 4°C for 10h. After the centrifugation is completed, collect 500ml in the first tube, then use a 50ml centrifuge tube to collect, 50ml / tube, 25-27 tubes can be collected. Detect the anti...

Embodiment 3

[0067] Example 3 Evaluation of various indicators of the vaccine prepared from the purified virus liquid of Sabin strain poliomyelitis type I, type II and III

[0068] 1. Impurity content

[0069]The superchaotrope 1 and superchaotrope 2 of the Sabin strain poliomyelitis type I, type II, and type III obtained in Example 2, and the molecular sieve purification solution obtained by using a traditional process in the control test, are then subjected to ion exchange and ultrafiltration Concentration and inactivation and other purification steps to obtain a monovalent virus stock solution. The obtained monovalent stock solutions of various types were diluted and mixed in equal proportions according to the antigen content in the finished product. The antigen content of the final product was 15 DU for type I, 45 DU for type II, and 45 DU for type III. Then according to the defined antigen content of the finished product, the finished product 1 (containing only effective antigenic co...

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Abstract

The invention provides inactivated vaccine produced through poliomyelitis attenuated strains and belongs to the field of preparation of biological products. According to the poliomyelitis inactivated vaccine, a traditional method that the inactivated vaccine is prepared from wild strains is changed, and the attenuated strains Sabin are adopted for producing the poliomyelitis inactivated vaccine. Due to the fact that I type virus liquid, II type virus liquid and III type virus liquid of the poliomyelitis strains Sabin are ultra-filtered, concentrated and subjected to sucrose density gradient centrifugation for ultra-filtration sucrose removing, sterilization and filtration, purified poliomyelitis virus liquid is obtained, solid virus particles and hollow virus particles in the poliomyelitis virus liquid can be effectively separated, impurity protein can be effectively removed, the content of the solid virus particles in centrifugal products is greatly increased, and the poliomyelitis inactivated vaccine which is good in immunogenicity, low in impurity content and high in safety and stability is prepared from the virus liquid.

Description

technical field [0001] The invention relates to the field of biological product preparation, in particular to an inactivated vaccine produced by using the attenuated poliovirus strain Sabin strain. Background technique [0002] Poliovirus belongs to the genus Enterovirus of the Picornaviridae family and contains a single-stranded positive-strand RNA gene. The protein capsid has an icosahedral stereosymmetric structure without an envelope. There are three serotypes of poliovirus, namely types I, II and III. Poliovirus particles include solid-structure virus particles and hollow-structure virus particles, and the solid-structure virus particles can react with body cells to produce neutralizing antibodies. Poliomyelitis is a highly contagious disease caused by the poliovirus, which mainly affects children under the age of five. Irreversible paralysis of the legs develops in 1 in 200 patients, and in more severe cases it causes difficulty swallowing and speaking. Respiratory ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/13A61P31/14
CPCY02A50/30
Inventor 姜德玉王一丁姚志东佟伟张星星沈建锋薛峰高强尹卫东
Owner SINOVAC BIOTECH
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