TY - JOUR
T1 - Purification of GCT cell-derived human colony-stimulating factors
AU - Erickson-Miller, C. L.
AU - Abboud, C. N.
AU - Brennan, J. K.
PY - 1988
Y1 - 1988
N2 - We have purified human-active colony-stimulating factors from the human cell line, GCT, using sequential ultrafiltration; cation exchange, gel permeation, and reverse-phase high performance liquid chromatography (RHPLC); and ion-exchange HPLC. Activity eluted from sodium dodecyl sulfate-polyacrylamide gels with a peak at 17,500 daltons. Similar results were obtained by processing 35S-methionine-labeled conditioned medium, which showed a labeled band in the same region of activity. This purified HPLC fraction, which had a specific activity of greater than 1 x 107 colonies/mg protein, stimulated neutrophil colonies at day 7 and neutrophil, neutrophil-macrophage, and eosinophil colonies at day 14 of culture, suggesting that it contained both granulocyte (G-CSF) and granulocyte-macrophage (GM-CSF) colony-stimulating factors. It also promoted the growth of erythroid progenitors, and the GM-CSF fraction purified by hydrophobic chromatography had erythroid-enhancing activity. Separation of G-CSF from GM-CSF was accomplished by the addition of trifluoroacetic acid to the mobile phase at the reverse-phase HPLC step.
AB - We have purified human-active colony-stimulating factors from the human cell line, GCT, using sequential ultrafiltration; cation exchange, gel permeation, and reverse-phase high performance liquid chromatography (RHPLC); and ion-exchange HPLC. Activity eluted from sodium dodecyl sulfate-polyacrylamide gels with a peak at 17,500 daltons. Similar results were obtained by processing 35S-methionine-labeled conditioned medium, which showed a labeled band in the same region of activity. This purified HPLC fraction, which had a specific activity of greater than 1 x 107 colonies/mg protein, stimulated neutrophil colonies at day 7 and neutrophil, neutrophil-macrophage, and eosinophil colonies at day 14 of culture, suggesting that it contained both granulocyte (G-CSF) and granulocyte-macrophage (GM-CSF) colony-stimulating factors. It also promoted the growth of erythroid progenitors, and the GM-CSF fraction purified by hydrophobic chromatography had erythroid-enhancing activity. Separation of G-CSF from GM-CSF was accomplished by the addition of trifluoroacetic acid to the mobile phase at the reverse-phase HPLC step.
UR - http://www.scopus.com/inward/record.url?scp=0023891870&partnerID=8YFLogxK
M3 - Article
C2 - 2448161
AN - SCOPUS:0023891870
SN - 0301-472X
VL - 16
SP - 184
EP - 189
JO - Experimental Hematology
JF - Experimental Hematology
IS - 3
ER -