[Purification of recombinant erythropoietin by reversed-phase high performance liquid chromatography (RP-HPLC)].
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.
Zou
Z
Z
China Medical University, Shenyang, 110001.
Sun
K
K
Lou
D
D
Hu
M
M
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004921:Erythropoietin / Q000302:isolation & purification*; D011994:Recombinant Proteins
D011994:Recombinant Proteins; D004921:Erythropoietin