Expression And Purification Of Interleukin-35

Interleukin is an immune factor in human body which plays an important role for human physiological function. There are numerous members in family of Interleukins, while interleukins 35(IL-35) which belongs to interleukin 12 family is an important inhibitory cytokines composed by heterologous dimers. Interleukin 35 plays an important role in immune regulation, involving in thoroughly cure of many kinds of inflammation, related with the body’s immune tolerance, treatment of autoimmune diseases and many other aspects. Base on current reports, IL-35 is effective for many diseases such as cancer, EAE, inflammatory and arthritis. In cancer area, for instance, it is able to increase the sensitivity of cancer cells apoptosis rate and can inhibit the proliferation of cancer cell simultaneously, which may be a breakthrough in this area. In addition to cancer, many autoimmune diseases are caused by the excessive immunity. IL- 35 can inhibit the proliferation of T cells, in the meantime lower immune response, which is a hotspot research area currently. As a latest inhibitory cytokines, it is necessary to have a more extensive study on IL-35.In this dissertation, the study mainly focus on protein expression and purification of IL-35 and receptor. Multiple clonal expression vectors were built and clonal expression vector of IL-35 with two sub P35, EBi3 and gp130 were constructed successfully for stable expressing. In addition, clean protein with good behavior was prepared by a suitable purified method. Many means of expression were achieved successfully such as eukaryotic expression, prokaryotic expression and inclusion body expression, and produce protein. These experiments laid the foundation for follow-up study.