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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.
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Article info
Publication history
Accepted:
October 9,
1968
Received:
May 20,
1968
Footnotes
☆Supported in part by Grants HE 10591-04(HEM) and MO-1-FR-61-05, from the National Institutes of Health.
☆☆The material in this paper was presented in part at the meeting of the American Association for the Advancement of Science, December 27, 1967, and the XII Congress, International Society of Hematology, September 5, 1968, New York, N. Y.
Identification
Copyright
© 1969 Published by Elsevier Inc.