Blood, 1960, Vol. 16, No. 4, pp. 1398-1410.
© 1960 American Society of Hematology, Inc.
Separation and Properties of Urinary Hemopoietine
G. HODGSON ,
S. FISCHER ,
M. PERRETTA ,
I. ESKUCHE ,
G. ARAYA , and
M. DINAMARCA
Methods are described for the separation of nondialyzable material with
erythropoietic activity (urinary hemopoietine), from urine of rabbits made
anemic (< 5g Hb/100) by phenylhydrazine and of patients with aplastic
anemia (< 5g Hb/100). The material obtained was assayed for its effects on
Fe59 distribution 3 and 24 hours after tracer injection. The active extracts
produced a marked increase in the rate of Fe59 clearance from plasma, and
in the fraction of the injected Fe59 appearing in erythrocytes at 3 and 24
hours. Effect of the extracts was seen to be a linear function of log dose. The
extracts prepared from urine of patients with aplastic anemia had a specific
activity about 3 times higher than that obtained from urine of phenylhydrazine
rabbits. The fractions of highest specific activity obtained from rabbit’s urine
were (a) that precipitating between 50-75 per cent ethanol at pH 4.5; (b)
adsorbed by kaolin at pH 4.5 and eluted at pH 7-8; and (c) adsorbed by
D.E.A.E. cellulose at pH 4.5 and eluted by 0.2 M Na2HPO4 in 0.5 M. NaC1.
Only the ethanol procedure was used for human urine, and it was seen that
specific activities of the 0-50 and 50-75 per cent fractions were similar.
The fractions obtained from human and rabbit urine showed great variability in hexose, sialic acid and hexosamine contents, and no clear correlation was apparent between gross chemical composition and erythropoietic
activity. Paper electrophoresis showed the presence of a single component
moving behind albumin, in both active and inactive extracts of human urine.
Submitted on December 14, 1959
Accepted on May 31, 1960
