Gene cloning, product preparation and use of Chinese interleukin-18 subtype (53 Arg IL-18)


Application Number: 00107993
Application Date: 2000.06.07
Publication Number: 1326992
Publication Date: 2001.12.19
Priority Information:
International: A61K38/20;A61P35/00;C07K14/54;C12N15/63
Applicant(s) Name: Shuanglu Pharmaceutical Co., Ltd., Beijing
Address:
Inventor(s) Name: Xu Mingbo;Wang Yongbo;Huang Xiangdong
Patent Agency Code: 00000
Patent Agent:
Abstract The present invention relates to the preparation of human recombination human interleuki-18. PCR cloned human 53Arg IL-18 cDNA segment and protocaryon expression vector, such as pBV220, are assembled into high efficiency expression plamid pIL-18, which is used to convert coil bacillus before temperature induction IL-18 gene expression, with the expression amount reaching to 30-40 % of total thalli protein. Engineering bacteria are fermentation cloned anc crushed to extract inclusion body, the inclusion body is purified and urea denatured, and the product is two-step purified through gel exclusion chromatography and anionic exchange chromatography. Between the two steps, the product is molecular renaturated and added with protective agent to product freeze dried preparation, which may be used in the treatment of cancer and other disease.