Method of preparing vaccine through suspension culture of mammal cells and application of the method

A technology of cell suspension and cell culture, applied in biochemical equipment and methods, microorganisms, pharmaceutical formulations, etc., can solve problems such as high price, high cost of antigen production, difficulty in cell digestion and dissociation, and achieve production cost reduction, virus The effect of increasing content and reducing dependence

Active Publication Date: 2016-02-10
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this application uses an expensive sheet carrier, and the cost of antigen production is high; at the same time, after using a sheet carrier, it is difficult for cells to digest and dissociate from the carrier, and continuous amplification cannot be achieved

Method used

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  • Method of preparing vaccine through suspension culture of mammal cells and application of the method
  • Method of preparing vaccine through suspension culture of mammal cells and application of the method
  • Method of preparing vaccine through suspension culture of mammal cells and application of the method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1, utilize bioreactor microencapsulation PK15 cell to produce PCV2 vaccine

[0030] (1) Cell preparation: After digestion of PK15 cells in the logarithmic growth phase, adjust the cell density to 2×10 6 cell / ml is used as coating material;

[0031] (2) Preparation of capsule material solution: add sodium alginate into ultrapure water according to the final concentration ratio of 15g / L, stir evenly, sterilize at 115°C for 30min, and set aside;

[0032] (3) Preparation of aqueous phase mixture: according to light CaCO 3 and ultrapure water 0.3 parts: 2 parts mass volume ratio (g / ml) after stirring evenly, after sterilizing at 115°C for 30 minutes, take 2 parts (ml), add 20 parts of sterile calcium alginate solution prepared in step (2) (ml), after mixing evenly, add 22 parts (ml) of the PK15 cell seed solution prepared in step (1), so that the final concentration of PK15 cells is 1×10 6 cell / ml, stir well and use as water phase;

[0033](4) Preparation of t...

Embodiment 2

[0046] Example 2, Evaluation of the immune effect of PCV2 vaccine produced by microencapsulated PK15 cells in a bioreactor

[0047] Using the self-made PCV2 inactivated vaccine in Example 1 of the present invention, the immunization procedure is as follows: firstly, 7-day-old piglets are immunized for the first time, and 21 days later, the piglets are immunized for the second time. At the same time of immunization, the porcine PCV2 vaccine that has been marketed was immunized in the same way to evaluate the difference in immune effect between the PCV2 inactivated vaccine prepared by microencapsulation technology and the marketed vaccine.

[0048] 1. Materials

[0049] PCV-2 single-component vaccine, prepared according to Embodiment 1, wherein the content of each part of virus liquid ≥ 1 × 10 6.5 TCID 50 .

[0050] Porcine circovirus type 2 inactivated vaccine, Pulaike Bioengineering Co., Ltd., batch number 20130802.

[0051] 2. Test method

[0052] 20 double-negative pigl...

Embodiment 3

[0059] Embodiment 3, utilize bioreactor microencapsulation ST cell to produce classical swine fever virus vaccine

[0060] (1) Cell preparation: After digestion of ST cells in the logarithmic growth phase, adjust the cell density to 2×10 6 cell / ml is used as coating material;

[0061] (2) Preparation of capsule material solution: add sodium alginate into ultrapure water according to the final concentration ratio of 15g / L, stir evenly, sterilize at 115°C for 30min, and set aside;

[0062] (3) Preparation of aqueous phase mixture: according to light CaCO 3 and 0.3 parts of ultrapure water: 2 parts of weight-volume ratio (g / ml), after stirring evenly, after sterilizing at 115°C for 30 minutes, take 2 parts (ml), and add 20 parts of sterile calcium alginate solution prepared in step (2) (ml), after mixing evenly, add 22 parts (ml) of ST cell seed solution prepared in step (1), so that the final concentration of ST cells is 1×10 6 cell / ml, stir well and use as water phase;

[0...

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Abstract

The invention provides a method of preparing a vaccine through suspension culture of mammal cells, which includes the following steps: (1) preparing microencapsulated animal cells through a microencapsulation method; (2) inoculating the microencapsulated animal cells in a cell culture system to perform cultivation; and (3) inoculating virus into the cell culture system to culture the virus, and obtaining a virus liquid to prepare the vaccine. By means of the microencapsulation method to culture the animal cells, a problem of large-scale continuous amplification of cells is solved. When the cultured animal cells are used for breeding virus, a high content of virus is achieved, so that the virus liquid, when being used for preparing the viral vaccine, can also achieve a better immune effect. The invention also provides a method of preparing a cell-expression product through the suspension culture of the mammal cells. The method reduces dependency on micro-carriers and is greatly reduced in production cost.

Description

technical field [0001] The invention belongs to the field of biological products. Background technique [0002] One of the most exciting developments in cell culture is the use of microcarriers (MC). The carrier, first reported by Van Wezel, has been improved and expanded in scope after extensive work by a research group at the Massachusetts Institute of Technology (MIT). At present, there are more than ten kinds of commodity MCs sold in the international market. Currently commonly used commercial microcarriers include cytodex series, biosilon series, solo microcarriers, Cultipher microcarriers, and Fibradisk sheet carriers. Large, not suitable for industrial production. [0003] Microencapsulation technology is mainly used for microencapsulation of viral vaccines and chemical drugs, through the sustained release of microcapsules, so as to improve the prevention or treatment effect of test animals. The application of microencapsulation culture in vaccine development main...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/295C12N7/00A61P31/14A61P31/20
Inventor 张许科孙进忠王进产田克恭
Owner PU LIKE BIO ENG
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