Fermentation, Purification and Identification of Recombinant Human Interleukin-18

VernacularTitle:重组人白介素18的发酵、纯化及功能鉴定
Author: Xiumei AN ; Lili YANG ; Hui LI ; Shui CAO ; Xiubao REN
Publication Type:Journal Article
Keywords: interleukin-18; pichia; fermentation; culture media; isolation & purification
From: Tianjin Medical Journal 2009;37(7):559-561,后插4
CountryChina
Language:Chinese
Abstract: Objective: To investigate the expression of recombinant human intedeukin-18 (hIL-18) in 3.7 L fermenter with the constructed engineer train Pichia pastoris X-33/hIL-18, and the procedures of expression and purification thereof.Methods: The train X-33/hIL-18 was inoculated in BMGY medium and then inoculated into the fermenter until the A600 of the culture reached about 6. The supernatant of fermentation was isolated and purified with centrifugal fiher devices, hydrophobic chromatography column and anion exchange chromatography column. Results: The recombinant hIL-18 was expressed in 3.7 L fermenter with batch feed methanol and the concentration reached 202 mg/L. After the purification, the purity could be more than 97%. The recombinant hIL-18 was shown to induce interferon-gamma (IFN-γ) production by human peripheral blood mononuclear cells (PBMCs), and enhanced NK cell cytotoxicity synergistically with IL-2. Conclusion: A great deal of the recombinant hIL-18 with higher purity could be harvested by Pichiapastoris expression system. This study showed a new potential for further study of its function and activities.