Preparation of human erythropoietin for tissue culture.
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.
D002850:Chromatography, Gel; D046508:Culture Techniques; D004355:Drug Stability; D004921:Erythropoietin / Q000302:isolation & purification* / Q000652:urine; D006801:Humans; D008722:Methods
D004921:Erythropoietin