Methods for the purification, assay, characterization and target cell binding of a colony stimulating factor (CSF-1)

Separation of functionally distinct human granulocyte-macrophage colony-stimulating factors.

Colony Stimulating Factors

The fractionation, characterization, and subcellular localization of colony-stimulating activities released by the human monocyte-like cell line, GCT.

A method for purifying CSF protein is described. The method comprises: precipitating the protein with ammonium sulfate at 80% saturation to form a pellet containing the CSF protein; resuspending the pellet in a buffered solution at a pH in the range of about 6 to about 8; applying the buffered solution containing CSF to a chromatographic column, eluting with the buffered solution containing sodium chloride and collecting the fractions having CSF activity; pooling the active fractions, applying them to a C4 reverse phase column and eluting with a 0 to 90% acetonitrile gradient to collect the active fractions. The purified CSF protein has a specific activity of at least about 1×10 7 units per mg of protein and preferably at least about 4×10 7 units per mg of protein when assayed using the human bone marrow assay.

Isolated native primate GM-CSF protein

https://febs.onlinelibrary.wiley.com/doi/pdf/10.1016/0014-5793%2878%2980385-8

Yvonne Purcell

Papers

Purification of a colony stimulating factor from culture cell lines propagated from human lung.