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  • Title: Preparation of human erythropoietin for tissue culture.
    Author: Cutler RL, Johnson GR, Nicola NA.
    Journal: Exp Hematol; 1985 Sep; 13(8):796-801. PubMed ID: 4043260.
    Abstract:
    A simple two-step procedure has been developed for the production of human urinary erythropoietin suitable for tissue culture use. Lyophilized human urinary protein was fractionated by phenyl-Sepharose chromatography, the erythropoietin binding in 2 M lithium chloride and being eluted by an ascending linear gradient to 6 M guanidine hydrochloride. The erythropoietin was further purified by ethanol (75% vol/vol) precipitation. This two-step procedure resulted in a 25-fold purification of erythropoietin with a 50% recovery of the starting activity. The erythropoietin preparation was suitable for use in murine or human cultures, being free of protease activity, inhibitory factors, colony-stimulating factors, and erythroid-enhancing activities.
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