| IFNα-2a is one of cytokine from human, which can restrain reproduction ofvirus involved RNA virus and DNA virus. Moreover, IFN α-2a can regulateimmune function and inhibit growth of tumor cytokine. Industrialization ofIFN could be made with the rapid development of biological technology.IFNα-2a made in china which was produced by Bodies Technology andprocaryon system at present. Process of technology and final productioncontain some shortages caused by feature of Bodies Technology, such as,recombination human IFN α-2a increased one Met at the end of N in aminoacid sequence and increased the probability of antibody produced in humanbody compared to natural IFNα-2a. The course of degeneration andrenaturation is complex and then it effect the efficacy of Purification indownstream and denaturant effect greatly biological activity of the production.Human serum albumin was used as protectant which increased the chance ofgetting other diseases.Therefore, technology of recombination human IFN α-2a secreting from E.coli was studied based on construction of engineering bacteria. The main workis as followed:Prescription of fermentation medium was selected by study of laboratoryand semi-works production. The effect of adding singly carbon source andcarbon/nitrogen source on fermentation was compared then stabiletechnology of semi-works production of fermentation was gained.Purification technology was obtained by study of semi-works productionand amino acid sequence of stock solution gained by the technology accordswith that of natural amino acid. Purity of RP—HPLC and SEC—HPLC,concentration of bacterial .endotoxin, residual volume of host protein andDNA gained by the technology were higher than those in " ChinesePharmacopeia "Rational prescription of praeparatum was selected and curve of freeze-drytechnology was determined in this study. Stability of praeparatum was alsostudied. The results showed that : IFN α-2a produced by technology ofsecreting type expression could be preserved 2 months,6 month and 24months at 40±2℃,25±2℃ and 2～8℃ respectively. So, expiration datewas decided 2 years at 2～8℃. Result of stability experiment alsodemonstrated that freeze-dry technology we adopted is fit for IFN α-2a. Inaddition, latent danger of use was decreased because Human serum albuminwas not adopted as protectant in prescription of praeparatum.Stability of transfer of culture for engineering bacteriaEngineering bacteria for IFN α-2a /w3110 remained heredity stability after50 generations transfer on slope and 30 generations on flat.Above all, fermentation ,purification ,prescription of praeparatum andfreeze-dry technology for IFN α-2a /w3110 we studied are suitable and thestudy provided theory foundation for recombination of human IFN α-2a inchina.
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