Serum (newborn calf, fetal calf, swine fever)
||Should provide the drug production license or drug business license, product registration certificate, GMP certificate, production approval number of the material.
Quality standards of serum
Faint yellow, transparent and slight viscous liquid without hemolysis or foreign body.
Should be free of bacterial growth
Should be free of mycoplasma growth
[Exogenous virus test]
1. Cell inoculation culture: add the tested serum to the MEM culture medium at a concentration of 10%, inoculate VERO, BHK21, PK15 and the bovine testis cell monolayer cultured by the 48-well cell culture plate, each cell should be inoculated with 8 wells, set up 2 bovine viral diarrhea / mucosal disease virus control wells, culture at 37℃ for 5 days, observe the cytopathic effect, all the cells should not have cytopathic effects.
2. Fluorescent antibody test method: in the 5th day, use PBS to wash the 2 wells and virus control wells in each cell for three times, add 0.5 ml of 80% acetone to each well, fix below 4℃ for 30 minutes, abandon the acetone and dry naturally, add 0.3ml of BVDV fluorescent antibody staining to each well, dye at the room temperature for 30 minutes, abandon the fluorescent antibody and use PBS to wash for three times, observe with fluorescence microscopy, the tested serum well should be free of fluorescence staining.
3. Hemadsorption test method: for the remaining 6 cell wells, add 0.3ml of equivalent miscible liquids of 0.2% guinea pig erythrocyte, O-type human erythrocyte and chicken erythrocyte to each of the first three wells, effect at 2-8℃ for 30 minutes, add the same miscible liquids to the other three wells and effect at the room temperature for 30 minutes. Observe with microscope, wells should be free of hemadsorption.
[specific antibody test]
check by means of ELISA, should not contain antibodies such as BVDV, CSFV, PRRSV and TGEV.
Should be below 10EU/ml.
[cell proliferation test]
The absolute cloning efficiency of the reference serum should be no lower than 20%;
The absolute cloning efficiency of the fetal calf serum should be no lower than 80%;
The absolute cloning efficiency of the newborn calf serum should be no lower than 50%.
Process is verified to meet the requirements for production through process test.