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Process for preparing human diploid cell rabies vaccine through Celligen310 bioreactor

A technology of human diploid cells and bioreactors, applied in biochemical equipment and methods, microorganisms, antiviral agents, etc., can solve problems such as tumor risk, fever, and allergic reactions, and achieve maximum safety and preparation technology The effect of simple and easy industrial production

Inactive Publication Date: 2013-05-08
万里明
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These vaccines generally have local allergic reactions, sometimes systemic side effects such as fever, and even potential tumor-causing risks

Method used

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  • Process for preparing human diploid cell rabies vaccine through Celligen310 bioreactor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Embodiment 2: PM strain virus seed preparation

Embodiment 2

[0013] The PM strain of rabies virus was purchased from the Wistar Institute in the United States. According to the requirements of vaccine preparation, firstly, a PM-adapted virus strain adapted to human diploid cells is established.

[0014] The cells passaged by the above method are inoculated with the PM strain, the MOI is 0.01-0.001, the pH is adjusted to 7.6-7.8, and then cultured at 33°C-35°C. Harvest the virus after 96-120 hours, take a sample, and check the virus titer with the mouse inoculation method, and its lgLD 50 / mL is ≥ 7.0.

[0015] Identification test

[0016] The specificity of the virus species was identified by the mouse brain neutralization test. Make 10-fold serial dilution of the virus seed, mix the appropriate dilution of the virus solution with the same amount of rabies virus-specific immune serum, and set up a normal serum control group at the same time, and inoculate 11-13g mice at each dilution of the test group and the control group Inoculate...

Embodiment 3

[0024] When the cells were confluent, the cell surface was washed with phosphate buffered saline (PBS). Then replace it with a new culture medium, and adjust the pH of the culture medium to 7.6-7.8.

[0025] Example 4: Inoculation of virus

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Abstract

The invention belongs to the field of biological products, in particular to a process for preparing rabies vaccine by culturing human diploid cells WI-38 and MRC-5 (called as human diploid cells as follows) as toxigenic cells through a Celligen310 bioreactor and taking a rabies virus PM strain as a virus seed, wherein the finished vaccine is prepared by comprising the steps of resuscitating human diploid cells, culturing and proliferating the human diploid cells, inoculating the virus seed of the rabies virus PM strain onto the human diploid cells, domesticating, inoculating and culturing virus seeds, collecting virus filtrate, inactivating, purifying, concentrating, adding a protective solution and the like. Verification indexes of the vaccine meet standards of Chinese Pharmacopoeia of 2010 version. The process is characterized in that the human diploid cells are cultured and prepared as matrix cells by using the Celligen310 bioreactor; therefore, exogenous pollution factors and tumorigenicity of animal passage cells can be avoided. As the inoculated virus strain is the rabies virus PM strain in the preparation process of the vaccine, the immune effect is better than that of the traditional vaccine. The vaccine prepared by using the process has the advantages of being high in purity, good in immune effect and high in safety.

Description

1. Field of invention [0001] The invention belongs to the field of biotechnology, and in particular relates to a process for preparing human diploid cell rabies vaccine in a Celligen310) bioreactor. 2. Background of the invention [0002] Rabies is a zoonotic infectious disease caused by rabies virus, which has the highest fatality rate in the world. It is usually infected after being bitten by a virus-carrying animal. Once onset, the mortality rate is 100%. Rabies is widely distributed in the world, and people of different races are generally susceptible to rabies. The epidemic areas are mainly in Asia, Africa and Latin America. my country has the second highest incidence rate in the world after India. In recent years, the epidemic of rabies in my country has been on the rise, and the number of deaths ranks first among the 37 legally notifiable infectious diseases in my country. Fortunately, proper inoculation of rabies vaccine has a nearly 100% protection rate against r...

Claims

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Application Information

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IPC IPC(8): A61K39/205A61K9/19A61P31/14C12N7/00
Inventor 刘孝民刘培生
Owner 万里明
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