Dialysis as a tool in the study of erythropoietin

Jasper P. Lewis, Dorothy A. Alford, Russell R. Moores, Edward Gardner, Claude Starr Wright, Linda L. Smith, W. Arnold Scharnitzky, Willie A. Neal

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)154-162
Number of pages9
JournalThe Journal of Laboratory and Clinical Medicine
Volume73
Issue number1
StatePublished - Jan 1969
Externally publishedYes

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

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