Preparation And Quality System Of Canine Interferon Alpha

In recent years, dogs as pets have become more and more important in human lives. But various virus diseases disturb canine husbandry. Interferon is the natural protein and producted by a variety of cell virus infection or other biological induction agents stimulus. The main function of the interferon is the antiviral, immune activation and cell growth regulation. Canine interferon has broad-spectrum antiviral activity.Then it can treat canine distemper, canine parvovirus, canine viral hepatitis and other high fatal viral disease, and adjuvant cancer treatment. So design and synthesis of canine interferon has a very good application prospect.After codon optimization, the CaIFN-α gene fragments was synthesized, cloned into pBV220vector and expressed in E.coli BL21. The molecular weight of CaIFN-α was18KD and the expression of CaIFN-α was25%.Then the inclusion denaturated and renaturated, the purity was69%, and recovery rate reached28%. Hydrophobic interaction chromatography was used to purify the protein, the purity was87%, and recovery rate reached81%. Then the protein was purified by molecular sieving chromatography, the purity was96%, and recovery rate reached90%. The specific activity of CaIFN-α was3×10’IU/mg, the endotoxin was≤10EU/mg, and yield was20%. This process for production was suitable for large-scale production.