A simple three-step purification procedure for interleukin 3 involving absorption to fixed cells.

摘要: We have found that treatment of B6SUtA1 cells with 0.01% glutaraldehyde transformed them into mechanically resistant spheres, thereby making it possible to use these high interleukin 3 (IL-3) receptor-bearing as a solid phase reagent suitable for the large scale purification murine IL-3 (mIL-3). Using this technique, mIL-3 was purified from serum-free pokeweed mitogen-stimulated spleen cell-conditioned medium (PWM-SCCM) approximately 16,000-fold using absorption cells, Sephadex G75 superfine chromatography, and reverse performance liquid chromatography on C18 column. The overall yield 16%. final product consisted two proteins molecular weights 19.5 16.5 kd. Both species possessed mIL-3-like activity. N-glycanase preparation converted all 19.5-kd material lower weight species, suggesting represented different glycosylated states produced by activated T cells. This confirmed competition studies showed excess pure Escherichia coli-derived recombinant mIL-3, but not granulocyte-macrophage colony-stimulating factor (GM-CSF), could prevent binding both PWM-SCCM-derived