| Cell factory is polystyrene(PS) as material, using the static culture, in the limited space makes full use of cultured on the surface of a cell culture device, has been more and more applied to vaccines and other production preparation fields of biological products. Cell factory is a one-time use in foreign countries, but are repeated use in the country.Taking the vaccine quality and production costs into account, this has become the key to the application of cell factory in vaccine production. At present, there are mainly two kinds of high-energy radiation used for radiation sterilization: one is the radioactive isotope 60Co-gamma rays, the other is the high-energy electron beam generated by the electron accelerator. The research of gamma rays in irradiation sterilization was earlier, and the application is also more extensive.In this paper,the cell factory was used as materials.Different doses of 60Co-gamma rays and electron beam of the irradiation sterilization were done to cultivate 2BS cells, attenuated strains and its producing virus harvested in liquid cultures of HAV L-A-1.Through different indicators,a suitable dose for sterilization and using times were selected.Without affecting the product quality, it is proved that the sterilization effect was in line with the GMP requirements, reduced the 60 Co cell factories on the material damage,and increased the number of times using cell factories. It achieved energy-saving emission reduction, cost decreasing and benefit increasing.The company’s cost was reduced to the greatest. The main conclusions were as follows:1. Initial use of cell factory was applied to freeze dry hepatitis a reduction of live attenuated vaccine 2BS cell culture, HAV L-A-1 reduced virus culture and virus harvested liquid preparation.Under the microscope, cells were a dense layer of fibrous.Cell count, the titer of virus and bacterial endotoxin, mycoplasma, sterile and other indexes and finished products of bovine serum albumin residual amount, chloroform residual volume, abnormal toxicity test results were in line with the current edition of Chinese Pharmacopoeia and GMP requirements. Cell factory was further verified for freeze-dried hepatitis a reduction of live attenuated vaccine production preparation.2. Though the similarities and differences of different doses of 60Co-gamma rays and electron beam irradiation on the cell factory sterilization,15 kGy 60Co-gamma rays and electron beam can’t satisfy the requirements of sterilization on the cell factory; 20 kGy 60Co-gamma ray and electron beam can meet the 10 layer cell factory,and can be reused 6 times,but not meet the requirements for radiation sterilization of 40 layer cell factories for radiation sterilization requirements;25kGy,30 kGy, 35 kGy, a radiation dose of 60Co-gamma rays and electron beam can be effective to sterilize the cell factories,and to meet the aseptic GMP request.It was used to culture 2BS cells,and to prepare HAV L-A-1 attenuated virus and harvested virus liquid.From the cell counting, virus titer, sterile the bacterial endotoxin of Mycoplasma, bovine serum albumin, residual chloroform residue and abnormal toxicity test results.All meet the requirements of the Chinese Pharmacopoeia and GMP regulations.3. To study the different doses of 60Co-gamma rays and electron beam of Bacillus pumilus kill similarities and differences,20 kGy,25kGy,30 kGy,35kGy4 radiation dose 60Co-gamma rays and electron beam can be placed in the short spores from the surface of the cell factory 106cfu/ml and completely killed;25kGy,30 kGy,35kGy60Co-gamma rays and electron beam can be placed in the short spores from 106cfu/ml at the center of the cell factory and completely killed.They are able to effectively ensure the cell’s factory aseptic.4. The effects of different doses of 60Co-gamma rays and electron beam on the color, light transmittance, tensile strength, bending strength and pressure resistance of the cells were studied. The results showed that the irradiation sterilization had a certain effect on the microstructure of the cell factory. With the increase of radiation dose and the use of the number of times, the more serious damage to the cell factory.5. In summary,25 kGy was a optimal dose of 60Co-gamma rays and electron beam for sterilization,and cell factories in the fifth of hepatitis A vaccine product had no greater impact.Cell factory can meet the freeze dried attenuated for hepatitis A virus vaccine preparation conditions and guarantee no bacteria within six months after sterilization. |
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