Separation and Partial Characterization of Erythropoietin from Human Urine

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Abstract
Crude erythropoietically active fractions were obtained by precipitation of urine from anemic patients with 80 % acetone at pH 4.0. The precipitate was lyophilized, dialyzed, and again lyophilized. Results from chemical analysis and stability studies are presented for this crude urinary powder. The crude urinary powder (CUP) was purified further by the following procedure: 1) CUP was dissolved in 0.0375 M NaCl (pH 4.5) and introduced into a DEAE cellulose column, and the activity was eluted with 0.15 M NaCl containing 0.05 M Na2HPO4; 2) the DEAE eluent was adjusted to 0.001 M phosphate (pH 6.7) and introduced into a calcium phosphate column, and the activity was eluted with 0.005 M phosphate (pH 6.7); and 3) this final eluent was precipitated with 80% ammonium sulfate. This material contained as much as 136 units/mg of protein and the recovery was approximately 25%.

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