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 journalArticle

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 1 1969

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Medicine(all)

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    Lewis, J. P., Alford, D. A., Moores, R. R., Gardner, E., Wright, C. S., Smith, L. L., Scharnitzky, W. A., & Neal, W. A. (1969). Dialysis as a tool in the study of erythropoietin. The Journal of Laboratory and Clinical Medicine, 73(1), 154-162.