反相高效液相色谱法分离重组促红细胞生成素  

Purification of Recombinant Erythropoietin by Reversed Phase High Performance Liquid Chromatography(RP HPLC)

作　　者：邹钟诚[1,2] 孙开来[1,2] 娄丹 胡明 

机构地区：[1]中国医科大学 [2]沈阳三生公司

出　　处：《色谱》1998年第3期263-264,共2页Chinese Journal of Chromatography

摘　　要：利用反相高效液相色谱法对重组促红细胞生成素进行了纯化。结果表明，利用Ｃ４反相柱和乙腈－三氟乙酸流动相在洗脱梯度和上样样品纯度等条件都较适当时，可以简单、快速、高效地从粗品中分离出重组促红细胞生成素，获得的产品纯度高，接近１００％；比活性好，约为１．９６×１０８ＩＵ／ｇ蛋白。Recombinant erythropoietin is a glycoprotein which is strongly hydrophobic. We purified recombinant erythropoietin by RP HPLC. The results showed that 30nm Spherisorb C 4 reversed phase column and acetonitrile TFA mobile phase with gradient elution, could readily isolate recombinant erythropoietin from crude samples with high speed and efficiency. Erythropoietin could be eluted at the concentration of about 50% acetonitrile with appropriate sample purity and elution gradient. When RP HPLC is used as a final step of purification, the purity of sample should be at least 60%. The purity of the product tested by SDS PAGE was nearly 100%. And the specific activity of the procuct was about 1.96×10 8 IU/g protein.

关 键 词：促红细胞生成素 纯化 分离 重组EPO HPLC 

分 类 号：TQ464.7[化学工程—制药化工]