Research Article| Volume 73, ISSUE 1, P154-162, January 1969

Dialysis as a tool in the study of erythropoietin

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      This report re-examines the application of dialysis to the purification and characterization of erythropoietin. Dialysis was done on urinary concentrates obtained by column chromatography with a membrane that yielded an increase in the erythropoietin specific activity in the dialysate. Physicochemical and potentiation studies were done on the fractions obtained. By dialyzing against a fresh 0.5M NaCl solution each day, most of the dialyzable erythropoietic fraction was removed in 9 days leaving an appreciable amount of a nondialyzable erythropoietic factor. The sedimentation coefficients of the nondialyzable erythropoietic fraction ranged from 2.7S to 4.5S. The most highly purified dialyzable fraction had a sedimentation coefficient of 2S, contained 11 per cent protein, and by immunoelectrophoresis had 3 lines in the α-globulin-albumin region and one line in the γ-globulin region. Inhibition was demonstrated by increasing the amount of serum used. The Lineweaver-Burk plot of the data for the nondialyzable fraction resulted in a curve typical of enzymatic inhibition by excess substrate.
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