Purification of murine erythropoietin produced in serum-free cultures of erythroleukemia cells.
We previously documented that several erythroleukemia cell lines released factors that stimulated erythropoiesis in vivo and in vitro. A simple five-step scheme has been devised that allows purification of this erythropoietic activity to apparent homogeneity. The methods employed included lectin affinity chromatography (wheat germ agglutinin), gel filtration (ultro gel ACA44), ion exchange, hydroxylapatite, and high performance liquid chromatography. Following polyacrylamide gel electrophoresis, biologic activity was recovered in an area corresponding to a molecular weight of 35,000 daltons. Silver staining of a polyacrylamide gel after electrophoresis of our most purified preparation revealed a single band at 35,000 daltons.
Qian
R L
RL
Chin
K
K
Kim
J K
JK
Chin
H M
HM
Cone
J
J
Hankins
W D
WD
D000818:Animals; D001769:Blood; D002460:Cell Line; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004921:Erythropoietin / Q000096:biosynthesis / Q000302:isolation & purification*; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism*; D007942:Leukemia, Experimental / Q000378:metabolism*; D051379:Mice
D003470:Culture Media; D004921:Erythropoietin