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Mass-production method of hydrophobic vaccine

A rabies vaccine, a large-scale technology, applied in the direction of antiviral agents, pharmaceutical formulas, medical preparations containing active ingredients, etc., can solve the problems of small loading, only batch cultivation, high cost of equipment and supporting facilities, etc. Achieve simple and efficient separation and realize the effect of large-scale production

Inactive Publication Date: 2007-05-23
广州市嘉合生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the small amount in the bioreactor (3-5 g / L), it can only be used for batch cultivation, and the industrial scale requires a larger volume of bioreactor, generally up to 1000 liters, equipment and supporting facilities higher cost

Method used

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  • Mass-production method of hydrophobic vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] (1) First, the Vero cells were recovered, expanded and cultured in a spinner bottle, and then 3.5×10 5 The density of / ml was inoculated into the Celligen Plus bioreactor tank with fixed bed basket stirring system (capacity can be 2.2 liters, 5 liters, 7.5 liters or 14 liters) with Fibra-Cel disks as the carrier, and cultured with cell proliferation The culture medium for expansion was carried out, wherein the Fibra-Celdisks carrier was used in an amount of 30 grams per liter of tank volume, and the cell proliferation culture medium used was 199 medium plus 6% bovine serum and an appropriate amount of gentamicin sulfate. The conditions for cell proliferation culture were pH 7.2, temperature 37° C., dissolved oxygen (DO) 50%, and stirring speed 120 rpm.

[0043] (2) When the Vero cells grow to a certain density, generally 6-12 days, then replace the cell maintenance medium with a multiplicity of infection (MOI: the ratio of the number of viruses used for infection to the...

Embodiment 2

[0050] (1) First, the Vero cells were recovered, expanded and cultured in a spinner bottle, and then 5×10 5 The density of / ml is inoculated into the BioFlo 4500 bioreactor tank (capacity can be 20 liters or 30 liters) with a fixed bed basket stirring system with Fibra-Cel disks as the carrier, and the cell proliferation culture medium is used for expansion culture. The amount of Fibra-Cel disks carrier used is 40 grams per liter tank volume, and the cell proliferation culture medium used is 199 medium plus 2% bovine serum and an appropriate amount of gentamicin sulfate. The conditions for cell proliferation culture were pH 6.9, temperature 36° C., dissolved oxygen (DO) 30%, and stirring speed 30 rpm.

[0051] (2) When the Vero cells grow to a certain density, generally 6-12 days, then replace the cell maintenance medium with a multiplicity of infection (MOI: the ratio of the number of viruses used for infection to the number of cells) of 0.025-0.125 or the final Concentratio...

Embodiment 3

[0058] (1) First, the Vero cells were recovered, expanded and cultured in a spinner bottle, and then 25×10 5 The density of / ml was inoculated into the BioFlo Pro bioreactor tank with fixed bed basket stirring system (capacity can be 75 liters, 150 liters, 300 liters) with Fibra-Cel disks as the carrier, and the cell proliferation culture medium was used for expansion. For augmentation culture, the amount of Fibra-Cel disks carrier used is 30 grams per liter of tank volume, and the cell proliferation culture medium used is 199 medium plus 10% bovine serum and an appropriate amount of gentamicin sulfate. The conditions for cell proliferation culture were pH 7.4, temperature 38° C., dissolved oxygen (DO) 70%, and stirring speed 150 rpm.

[0059] (2) When the Vero cells grow to a certain density, generally 6-12 days, then replace the cell maintenance medium with a multiplicity of infection of 0.025-0.125 (MOI: the ratio of the number of viruses used for infection to the number of...

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Abstract

The invention relates to a method for producing batch hydrophobia vaccine, wherein it comprises that: (1), in the biological reactor with fixed bed mixing system and Fibra-Cel disks carrier, using cell increment cultivate liquid to cultivate Vero cell; (2), when the Vero cell grows to some density, using cell hold cultivate liquid, seeding hydrophobia vaccine, and affecting cell; (3), increasing virus; (4), obtaining virus continuously; (5), ultra-filter concentrating and inactivating via beta-propanolide; (6), using protamine sulfate or DNA enzyme treatment to remove Vero cell DNA; (7), using Sepharose 4 Fast Flow as stuff to process chromatography purification; (8), adding human blood albumin and sugar as protector to be made into liquid agent; or adding human blood albumin, sugar, and gelatin, as protector and shaping agent to be made into dried agent. The inventive method can produce virus continuously in small biological reactor, to realize batch production.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and in particular relates to a method for large-scale production of rabies vaccine. Background technique [0002] Rabies is a natural foci infectious disease transmitted by animals such as dogs, cats, pigs, mice, and bats. Once a person is bitten by such an animal carrying rabies virus, he will be infected with the disease, and the fatality rate is 100%. . According to incomplete statistics, there are nearly 30,000 cases of rabies in Asia every year. my country is a high-incidence area of ​​rabies. In the 1980s, more than 5,000 people died every year, and it declined slightly in the 1990s. However, the incidence of rabies in my country has an upward trend in recent years. Rabies has no special and effective treatment, and injection of rabies vaccine is the only effective means to control the prevalence of rabies. [0003] Since Louis Pasteur proved in 1885 that the spinal cord of rab...

Claims

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Application Information

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IPC IPC(8): A61K39/205A61P31/14
Inventor 刘建青何春辉吴雪红
Owner 广州市嘉合生物技术有限公司
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