Production method for hepatitis A virus for vaccine production
A technology for hepatitis A virus and vaccine production, applied in the biological field, can solve the problems of high production intensity, low production level, complicated operation, etc., and achieve the effects of improving yield and quality stability, low production intensity, and ensuring production quality
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Embodiment 1
[0026] A production method of hepatitis A virus for vaccine production, comprising the following steps in sequence:
[0027] Step 1: Recovery and subculture expansion of cells: recovery of cells, and culturing cells in a granular solid microsphere carrier bioreactor;
[0028] Step 2: inoculate hepatitis A virus seeds: when the cell concentration is 10 6 Individuals / ml, the culture medium was discarded, and the HM175 strain of hepatitis A virus was mixed and inoculated according to the ratio of 0.001 MOI;
[0029] Step 3: maintaining and culturing the cells infected with the HM175 strain of hepatitis A virus: maintaining and culturing the cells infected with the HM175 strain of hepatitis A virus with MEM maintenance solution, replacing the MEM maintenance solution every day, and maintaining the culture for 14 days;
[0030] Step 4: Detect the replication of virus infection: From the 14th day, samples were taken every day, and the virus antigen in the cells was measured by immu...
Embodiment 2
[0036] A production method of hepatitis A virus for vaccine production, comprising the following steps in turn:
[0037] Step 1: recovery and passage expansion of cells: recovery of cells, and cultivation of cells in a porous microsphere carrier bioreactor;
[0038] Step 2: inoculate hepatitis A virus seeds: when the cell concentration is 10 7 Individuals / ml, the culture medium was discarded, and the HM175 strain of hepatitis A virus was mixed and inoculated according to the ratio of 0.01 MOI;
[0039] Step 3: maintaining and culturing cells infected with HM175 strain of hepatitis A virus: maintaining and culturing cells infected with HM175 strain of hepatitis A virus with MEM maintenance solution, replacing MEM maintenance solution every day, and maintaining the culture for 28 days;
[0040] Step 4: Detect the replication of virus infection: From the 14th day, samples were taken every day, and the virus antigen in the cells was measured by immunofluorescence method to detect...
Embodiment 3
[0046] A production method of hepatitis A virus for vaccine production, comprising the following steps in sequence:
[0047] Step 1: Recovery and subculture expansion of cells: recovery of cells, and culturing cells in a granular solid microsphere carrier bioreactor;
[0048] Step 2: inoculate hepatitis A virus seeds: when the cell concentration is 10 6 Individuals / ml, the culture medium was discarded, and the HM175 strain of hepatitis A virus was mixed and inoculated according to the ratio of 0.001 MOI;
[0049] Step 3: maintaining and culturing the cells infected with the HM175 strain of hepatitis A virus: maintaining and culturing the cells infected with the HM175 strain of hepatitis A virus with MEM maintenance solution, replacing the MEM maintenance solution every day, and maintaining the culture for 14 days;
[0050] Step 4: Detect the replication of virus infection: From the 14th day, samples were taken every day, and the virus antigen in the cells was measured by immu...
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