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研究自构建的重组人白细胞介素－ ６（ｒｈＩＬ－６）基因工程菌株高效表达的影响因素及其ｒｈＩＬ－６产品中试工艺。方法：①将ｒｈＬ－６－ＰＢＶ重组质粒转化至ＲＲＩ、ＪＭ１０３和 ＤＨ５ ａ不同宿主菌中，在Ｍ９ＣＡ和 ＬＢ培养基中，４２℃诱导培养不同时间（３～６ ｈ），观测 ｒｈＩＬ－６不同表达效率。②采用 １０ Ｌ发酵罐诱导培养，经破菌－洗包－溶包初步纯化后，再经三步柱层析纯化，提取ｒｈＩＬ－６。结果：①ｒｈＩＬ－６在ＤＨ５ ａ大肠杆菌中（ＳＤＨ－９４５株）表达效率高；可达 ３９％，用 Ｍ９ＣＡ培基，４２℃诱导培养 ５ｈ可使表达效率提高到４１％，②ＳＤＨ－９４５菌株在５批 １０Ｌ Ｍ９ＣＡ发酵诱导培养 ５ ｈ，ｒｈＩＬ－６表达效率可达 ３５． ８７±２． ６５％。经初步纯化后，ｒｈＩＬ－６的纯度达 ７４． ８７±３． ６８％，再经三步柱层析后，纯度可达 ９５％以上，比活性达 ４ × １０８ Ｕ／ｍｇ，总得率稍低可达 ２３．１％．结论：①ＳＤＨ－９４５工程菌株在 Ｍ９ＣＡ培养基巾，４２℃诱导培养 ５ ｈ，ｒｈＩＬ－６表达效率最佳。②１０ Ｌ发酵和纯化工艺可获高效价、高比活性、高纯度的ｒｈＩＬ－６合格生物制品。
To study the factors that affect the high expression of recombinant human IL-6 genetically engineered strain and the pilot-scale process of rhIL-6 product. Methods: ① The rhL-6-PBV recombinant plasmid was transformed into different host strains of RRI, JM103 and DH5a. The cells were cultured at 42 ℃ for different time (3 ~ 6 h) in M9CA and LB medium. Express efficiency. ② The culture was induced by 10 L fermenter, and then the rhIL-6 was extracted by the three-step purification after preliminary purification by bacteria-washing-package-lysing. Results: ① The expression of rhIL-6 in E. coli DH5a (SDH-945 strain) was highly efficient (up to 39%). The efficiency of expressing DHIL-945 was increased to 41% with M9CA medium at 42 ℃ for 5 h. 5 batches of 10L M9CA fermentation induced 5 h, rhIL-6 expression efficiency of 35. 87 ± 2. 65%. After the initial purification, rhIL-6 purity of 74. 87 ± 3. 68%. After three-step column chromatography, the purity reached more than 95%, and the specific activity was 4 × 108 U / mg. The total yield was as low as 23.1%. Conclusion: (1) SDH-945 strain showed the best rhIL-6 expression in M9CA medium at 42 ℃ for 5 h. ② 10 L fermentation and purification process will be high titer, high specific activity, high purity rhIL-6 qualified biological products.