Study On The Isolation And Purification And Safety Of Recombinant Porcine Inte Rferonα To Pig

【Objective】 r Po IFN-α is a cytokine with a broad-spectrum antimicrobial effect,and immune function of the host and has significant application clinically. In order to make it can be from the laboratory to industrial production and clinical applications, optimization of r Po IFN-α industrial expression conditions and evaluation of its biosecurity are the key development sectors.【Method】 We used automatic fermentor to express r Po IFN-α, we used ammonium sulfate to precipitation r Po IFN-α,Then we used affinity chromatography to get high purity r Po IFN-α. Used the Marc-145 types of system to detect activity of r Po IFN-α.By double antibody sandwich ELISA r Po IFN-α with rabbit polyclonal and mouse monoclonal antibodies of two r Po IFN-α antibodies, C hoose 30 days of age and 2 piglets day age, muscle injection of different doses of r Po IFN alpha, using set up double antibodies sandwich ELISA to detect different time point piglets serum r Po IFN alpha concentration change, studies the metabolic rule of interferons in pigs. By animal toxicity test, gene residue detection and piglets to produce antibody test, etc., for safety evaluation of the genetic engineering r Po IFN alpha.【Results】 By the above method we obtained more than 98% purity of r Po IFN-α, and using Marc-145 / VSV system to measure its activity,which was up to 1.00 × 108 u / μL. r Po IFN-α maintaining time in vivo is positive correlation between the amount of injection, and is also positive correlation between pig’s age. No matter at any time after the injection, r Po IFN- content was 50mg/ dose group was significantly higher than that of the rest of the serum dose injection group(30 day old piglets, pig) in vivo r Po IFN- alpha maintain time reached 24 h, and the newborn piglets(2 day old pigs) in vivo metabolism time is only 3h. Acute toxicity test of mice, 10mg/ only the following dose does not produce any toxic effect on mice The r Po IFN-α produced by our laboratory had no gene residue,did not cause allergic reactions in pigs,and did not induce IFN antibodies. We did not find pathological changes of the heart, liver, lung, kidney, spleen, pancreas, lymph nodes and injection site muscle by pathological tissue sections.