Oral hexavalent reassorted rotavirus live vaccine

A rotavirus and live vaccine technology, applied in antiviral agents, viral antigen components, digestive system, etc., can solve the problems of small coverage and serotype coverage, and achieve the effect of facilitating operation

Inactive Publication Date: 2015-04-22
WUHAN INST OF BIOLOGICAL PROD CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The diversity of rotavirus strains determines that multivalent vaccines are the most effective way to prevent rotavirus diarrhea. However, the main rotavirus vaccines currently on the market, Lanzhou Institute of Biological Products Rotway and GSK Rotarix is ​​a monovalent serotype rotavirus vaccine, and the serotype coverage is small. Alt

Method used

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  • Oral hexavalent reassorted rotavirus live vaccine
  • Oral hexavalent reassorted rotavirus live vaccine
  • Oral hexavalent reassorted rotavirus live vaccine

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0045] Example 1. Preparation of virus stock solution of each serotype

[0046] Cell passage: The culture medium of each serotype virus is Vero cells. The working seeds of Vero cells are first resuscitated. After culturing for 3-5 days at 37°C, they are subcultured and expanded according to a certain ratio. The culture vessels are T25, T75, and T175 cells. Culture flasks, 10-layer cell factories, and 40-layer cell factories are cultured in 40-layer cell factories for 3-5 generations and then inoculated with G1, G2, G3, G4, G8, and G9 serotype rotavirus.

[0047] Virus culture: After the cell confluence in the cell factory reaches more than 90%, virus inoculation can be carried out. Before virus inoculation, select a cell factory for cell count. According to the count result, calculate the amount of virus required by MOI (multiplicity of infection) 0.005-0.2, and use trypsin to activate the virus. The activation conditions are 37°C, trypsin concentration 10-20μg / ml, and 15min-60min...

Example Embodiment

[0050] Example 2. Development of vaccine protective agent

[0051] (1) Determination of virus resistance to acid

[0052] Determine the lowest pH value that the hexavalent rotavirus vaccine can tolerate through experiments.

[0053] Mix the hexavalent serotype virus mixed vaccine (mix the hexavalent vaccine stock solution in equal volume) and 1% sodium citrate solution in equal volume.

[0054] (2) Prepare 7 disposable sterile 50ml centrifuge tubes, 6 of which are added with 2ml of hexavalent seedlings mixed in step (1), and the other one is added with 1ml of hexavalent mixed seedlings and 1ml DMEM as a control.

[0055] (3) Adjust the pH, add 0ml, 0.4ml, 0.6ml, 0.7ml, 0.8ml, 1.2ml of 0.1M hydrochloric acid to 6 centrifuge tubes, and the corresponding pH is 7.84, 5.02, 4.00, 3.50, 3.05, 2.00 .

[0056] (4) Place 7 centrifuge tubes in a 37℃ water bath and incubate for 2h.

[0057] (5) After the incubation time is over, add DMEM to 20ml immediately in each centrifuge tube.

[0058] (6) Disp...

Example Embodiment

[0086] Example 3. Screening of different formulations

[0087] The ingredients of the 5 vaccine protectants are shown in Table 4

[0088] Table 4 The content of each component of different protective agents

[0089]

[0090] Each formula was mixed with the most temperature-sensitive serotype G9 virus, and the accelerated stability test was carried out. The results are shown in Table 5

[0091] Table 5 Virus stability test of different protection formulas (all data are the average of three test results, unit: lgFFU / ml)

[0092]

[0093] It can be seen that the protection of the above five formulas for G9 meets the requirements. Among them, F5 is placed at 37 ℃ for 7 days, the G9 titer drop is the lowest, and the protection for G9 is the best, so F5 is the best rotavirus vaccine Protective formula.

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Abstract

The invention relates to an oral hexavalent reassorted rotavirus live vaccine. The oral hexavalent reassorted rotavirus live vaccine contains six main epidemic strain serotypes of G1, G2, G3, G4, G8 and G9, comprises 99.6% of G serotype viruses of group A bovine rotaviruses and has good effects of preventing and treating rotavirus-caused diarrhea. The invention provides the live vaccine comprising hexavalent rotavirus serotypes and used for oral inoculation. The oral hexavalent reassorted rotavirus live vaccine is used for preventing rotavirus-caused infant diarrhea and comprises 1, G1, G2, G3, G4, G8 and G9 serotype live vaccines and all the serotype stoste titers are in a range of 1-5*10<6>FFU/ml, and 2, the protection agent comprises 0.5-2g/L of citric acid, 50-150g/L of sodium citrate, 250-450g/L of cane sugar, 5-10mm of zinc chloride and 10-20mm of calcium chloride.

Description

technical field [0001] The invention belongs to the field of vaccine preparations, in particular to an oral hexavalent reassortment rotavirus live vaccine. Background technique [0002] Human rotavirus belongs to the genus Rotavirus of the Reoviridae family, and is a non-enveloped RNA virus with a particle diameter of about 75 nm and consists of three layers of icosahedral protein capsids. Its genome is a double-stranded RNA containing 11 segments, encoding 6 structural proteins (VP1, VP2, VP3, VP4, VP6, VP7) and 5 non-structural proteins (NSP1, NSP2, NSP3, NSP4, NSP5). The laminin is VP4 and VP7, the inner shell protein is VP6, and the nucleoprotein is VP1, VP2 and VP3. Genetic reassortment can occur between viruses of the same group. HRV can be divided into 7 groups (A~G) according to the different antigenicity of the viral inner coat protein VP6, and only three groups A, B and C cause human diarrhea. Group A is the most common cause of diarrhea in infants and young chi...

Claims

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Application Information

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IPC IPC(8): A61K39/15A61P31/14A61P1/12
Inventor 杨晓明徐葛林李庆亮何泗涛白萱姜志军程满荣张久威马涛董犇胡蓉刘涛梁婧陈金华姜礼朋冯冬扬杨彪林楠段凯杨邦玲徐晓曾凯
Owner WUHAN INST OF BIOLOGICAL PROD CO LTD
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