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Method for producing vaccine by virtue of culturing animal cells

A technology of animal cells and cultured cells, which is applied in the field of bioengineering and can solve problems such as low degree of automation, large differences in product batches, unstable quality of cells and vaccines, etc.

Inactive Publication Date: 2011-10-05
HANGZHOU AMPROTEIN BIOENG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the production of vaccines is mainly carried out through the spinning bottle process, but this method has a low degree of automation and high labor intensity, and because of the uncontrollable environment of the spinning bottle cell culture, the quality of the cells and vaccines is unstable, and the product batches vary greatly; On the other hand, the cell adsorption growth area provided by a single spinner bottle is limited, and the cell density can only reach 1-5×10 5 cells / ml, the production capacity is low, and at the same time, the only way to expand the production scale is to increase the number of bottles, resulting in greater demand for workshop scale, equipment access, personnel, etc., and greater labor.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1(10

[0027] Example 1 (Preparation of PRRS vaccine by culturing Marc-145 cells in a 10L bioreactor)

[0028] Porcine reproductive and respiratory syndrome virus: CH-1R strain.

[0029] After the sheet-shaped carrier is sterilized by gamma rays at 24 kGy, it is first soaked in PBS, then rinsed with PBS, soaked in culture medium overnight, and set aside.

[0030] Inoculate 1 × 10 on the sheet carrier in the bioreactor 7 A Marc-145 cell was cultured and grown at 30-60% DO, pH 7.0-7.2, and 37°C for 5-7 days, during which glucose digestion was detected, and the medium was changed according to the sugar consumption. , so that the amount of sugar can be maintained at a daily sugar consumption of more than 1g / L. With the increase of cell density, the daily sugar consumption will gradually increase until it reaches more than 2.5g / L.

[0031] Drain the culture medium, wash it twice with sterile PBS, and inoculate porcine reproductive and respiratory syndrome virus according to the MOI of v...

Embodiment 2(10

[0036] Example 2 (Preparation of avian influenza vaccine by culturing MDCK cells in a 10L bioreactor)

[0037] After the sheet-shaped carrier is sterilized by gamma rays at 24 kGy, it is first soaked in PBS, then rinsed with PBS, and then soaked in culture medium overnight, and set aside.

[0038] Inoculate 0.5 × 10 on the sheet carrier in the bioreactor 7 For MDCK cells, the DO was controlled at 30-60%, the pH was maintained at 7.2-7.4, and the temperature was 37°C. Detect glucose digestion, and perform fluid replacement or rehydration according to the sugar consumption to keep the sugar level above 1g / L. Basically, the cell density reaches the maximum within 5-7 days.

[0039] Empty the culture medium, wash twice with sterile PBS, and inoculate H5N1 virus according to the MOI of virus infection at 0.01-0.1. All strains that are generally adaptable and sensitive to MDCK cells or other susceptible cells are applicable to the method of the present invention.

[0040] Continu...

Embodiment 3(10

[0044] Embodiment 3 (10L bioreactor cultivates ST cell to prepare swine fever vaccine)

[0045] After the sheet-shaped carrier is sterilized by gamma rays at 24 kGy, it is first soaked in PBS, then rinsed with PBS, soaked in culture medium overnight, and set aside.

[0046] Inoculate 2 × 10 on the sheet carrier in the bioreactor 7 For ST cells, the DO was controlled at 30-60%, the pH was maintained at 7.0-7.2, and the temperature was 37°C. Glucose digestion was detected, and the medium was changed every 2-3 days according to the sugar consumption, so that the sugar content was maintained above 1g / L, and the maximum cell density was reached in 5-7 days.

[0047] Drain all the culture medium, wash the sheet carrier and cells twice with sterile PBS, and inoculate ST cells with a concentration of 2-3% spleen toxin. The methods of the present invention are applicable to all strains that are generally adaptive and sensitive to ST cells or other susceptible cells.

[0048] The ST ...

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PUM

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Abstract

The invention belongs to the field of vaccine preparation, and provides a method for producing a vaccine by virtue of culturing animal cells in a flake carrier. The method comprises the following steps: culturing vaccine stromal cells on the flake carrier; inoculating virus based on a certain concentration; continuously culturing cells and virus under a bioreactor control condition to obtain a virus solution; repeatedly freezing and thawing the flake virus and infection cells; and mixing all collected solutions to synthesize a virus suspension so as to prepare the vaccine. In the method, a flake carrier culturing technology is applied to producing the vaccine by virtue of culturing the animal cells in the bioreactor, the flake carrier provides a three-dimensional space and a larger specific surface area for cell growth and virus breeding, the cell density of the cells is higher, and the bioreactor controls culture conditions and other parameters to provide the best producing environment for cell growth and virus breeding, so that the virus titer is greatly reduced, and the vaccine yield and quality are more stable.

Description

technical field [0001] The invention belongs to the field of bioengineering, and specifically relates to a method for cultivating animal cells to produce vaccines, more specifically to a method for cultivating animal stromal cells to produce vaccines through sheet-like carriers. Background technique [0002] Traditional vaccine production technology often isolates and prepares virus liquid by inoculating chicken embryos or animals. However, due to individual and species differences, it is difficult for many viruses to find suitable sensitive animals, and the results are affected due to large individual differences. determination. With the development of cell culture technology, most viruses can have corresponding sensitive cells, which creates conditions for the proliferation of viruses. With the development of large-scale cell culture technology, as long as there are sensitive cells, large-scale Vaccine production. [0003] At present, the production of vaccines is mainly...

Claims

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Application Information

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IPC IPC(8): A61K39/00C12N7/00C12N5/071
Inventor 惠觅宙李峦峰杜春玲宋羚羚戚西京
Owner HANGZHOU AMPROTEIN BIOENG
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