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目的研究用于分离纯化假单胞菌表达重组干扰素α-2b蛋白的离子交换树脂。方法利用工程菌假单胞菌发酵获得的菌体进行裂解,经离心、沉淀、盐析、疏水层析、超滤、透析等步骤,获得重组人干扰素α-2b蛋白溶液,通过对比8种商品化离子交换树脂(DE 52、DEAE FF、Fractogel DEAE、Bio-sep DEAE、CM 52、CM FF、Fractogel COO-、Bio-sep CM)的动态载量、洗脱条件及纯化后的重组人干扰素α-2b蛋白的活性回收率,采用安捷伦2100生物分析仪比较纯化后的重组人干扰素α-2b纯度,考察离子交换树脂用于假单胞菌表达重组干扰素α-2b蛋白分离纯化的适用性。结果 FractogelDEAE和Fractogel COO-的动态载量最高,分别为183.04 mg/mL和107.00 mg/mL;Fractogel DEAE和CM 52洗脱效果较好;DEAE FF和CM FF纯化后的干扰素α-2b蛋白活性回收率较高,分别为82.38%和49.46%。结论 DEAE FF和CM FF适用于假单胞菌重组干扰素α-2b蛋白的分离纯化。
OBJECTIVE To study the ion exchange resin used to isolate and purify Pseudomonas for recombinant interferon alpha-2b protein. Methods The bacterial cells obtained from Pseudomonas aeruginosa fermentation were lysed and the recombinant human interferon alpha-2b protein solution was obtained by centrifugation, precipitation, salting-out, hydrophobic chromatography, ultrafiltration and dialysis. Dynamic loading of commercial ion exchange resins (DE 52, DEAE FF, Fractogel DEAE, Bio-sep DEAE, CM 52, CM FF, Fractogel COO-, Bio-sep CM), elution conditions and purified recombinant human interference The activity of α-2b protein was recovered. The purity of recombinant human interferon α-2b was compared with Agilent 2100 Bioanalyzer. The ion exchange resin was used to isolate and purify recombinant interferon α-2b protein expressed in Pseudomonas. applicability. RESULTS Fractogel DEAE and Fractogel COO- had the highest dynamic loadings of 183.04 mg / mL and 107.00 mg / mL, respectively. Fractogel DEAE and CM 52 showed better elution efficiencies. The purified interferon α-2b protein activity of DEAE FF and CM FF The recoveries were higher at 82.38% and 49.46% respectively. Conclusion DEAE FF and CM FF are suitable for the isolation and purification of Pseudomonas aeruginosa recombinant interferon alpha-2b protein.