Purification of human erythropoietin.

TLDR

The nature of the difference between these two components is not yet understood. The authors purified erythropoietin via a seven‑step protocol involving ion exchange chromatography, ethanol precipitation, gel filtration, and adsorption chromatography, achieving a 21 % yield and 70 400 units/mg potency. Human erythropoietin was purified to apparent homogeneity, yielding a 21 % product with 70 400 units/mg potency, a purification factor of 930, and a single electrophoretic component, though two fractions of identical potency and size but slightly different mobility at pH 9 were observed.

Abstract

Human erythropoietin, derived from urine of patients with aplastic anemia, has been purified to apparent homogeneity. The seven-step procedure, which included ion exchange chromatography, ethanol precipitation, gel filtration, and adsorption chromatography, yielded a preparation with a potency of 70,400 units/mg of protein in 21% yield. This represents a purification factor of 930. The purified hormone has a single electrophoretic component in polyacrylamide gels at pH 9, in the presence of sodium dodecylsulfate at pH 7, and in the presence of Triton X-100 at pH 6. Two fractions of the same potency and molecular size, by sodium dodecyl sulfate gel electrophoresis, but differing slightly in mobility at pH 9, were obtained at the last step of fractionation. The nature of the difference between these two components is not yet understood.