Purification of Recombinant Erythropoietin by Reversed Phase High Performance Liquid Chromatography(RP HPLC)
反相高效液相色谱法分离重组促红细胞生成素

Recombinant erythropoietin is a glycoprotein which is strongly hydrophobic. We purified recombinant erythropoietin by RP-HPLC. The results showed that 30 nm Spherisorb C4 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 product was about 1.96 x 10(3) IU/g protein.