Purification to homogeneity of biologically active human interleukin 1.
Human interleukin 1 (IL-1) produced by lipopolysaccharide-stimulated monocytes was purified to homogeneity with retention of biological activity. IL-1 was measured by its ability to enhance the proliferative response of thymocytes to phytohemagglutinin. The purification procedure included hydrophobic affinity chromatography on phenyl Sepharose, gel filtration through Ultrogel AcA54 and preparative isoelectric focusing. Both charged species of IL-1, pI 5.1 and 6.8 have a molecular weight of 14,500 as determined by SDS-polyacrylamide gel electrophoresis. The complete purification resulted in a recovery of approximately 0.01% of IL-1 protein and if protection against losses by denaturation and adsorption in the final purification step was provided by bovine serum albumin, approximately 11% of IL-1 activity can be recovered.
D000818:Animals; D001681:Biological Assay; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007375:Interleukin-1 / Q000302:isolation & purification* / Q000502:physiology; D007525:Isoelectric Focusing; D051379:Mice; D009000:Monocytes / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism; D013936:Thymidine / Q000378:metabolism; D014462:Ultrafiltration
D007375:Interleukin-1; D013936:Thymidine