A NEW PREPARATIVE METHOD FOR ISOLATION OF HUMAN ERYTHROPOIETIN WITH HYDROPHOBIC INTERACTION CHROMATOGRAPHY

  • 1 January 1980
    • journal article
    • research article
    • Vol. 56  (4) , 620-624
Abstract
A new preparative method for isolation of human urinary erythropoietin (Ep) was developed using hydrophobic interaction chromatography on phenyl-Sepharose CL4B. Crude urine and urine concentrates from anemic patients were used directly without prior manipulation. In addition to facilitating hydrophobic interactions, phenyl-Sepharose provided .pi.-.pi. interactions between its phenyl group on the gel matrix and the aromatic amino acid residues of Ep, and contributed to specific resolution. Over 90% of the urinary contaminants were excluded from the column, and Ep was selectively bound. Its activity was eluted with 20% ethylene glycol in 10 mM NaOH containing 4 M guanidine hydrochloride. This single step offered a mean purification factor of 110 with a recovery of 85%.