Partial purification of guinea pig MIF by affinity column chromatography using macrophages.
First we have confirmed the previous observation that the macrophage migration inhibitory factor (MIF) was adsorbed on normal peritoneal macrophages when they were incubated at 4 C for 60 min. It was found that macrophages fixed with 2% glutaraldehyde gave more reproducible results than viable cells in terms of "adsorption" of guinea pig MIF. The adsorption was achieved more completely at 37 C than at 4 C, indicating that this reaction is a temperature-dependent phenomenon. Using these glutaraldehyde-fixed macrophages, a kind of cell-affinity column was successfully developed. The guinea pig MIF preparation lost its activity when it was passed through this affinity column, and MIF adsorbed on the column was recovered by elution with 0.1 M (L)-fucose of 0.1 M (D)-glucose. Such MIF active eluate was found to be at least 30--40 fold more pure than the original MIF preparation which had been previously fractionated according to its molecular weight. Therefore, this type of macrophage-affinity column may be useful for the purification of MIF.
D000327:Adsorption; D000818:Animals; D002846:Chromatography, Affinity; D005643:Fucose / Q000494:pharmacology; D005976:Glutaral / Q000494:pharmacology; D006168:Guinea Pigs; D008263:Macrophage Migration-Inhibitory Factors / Q000302:isolation & purification*; D008264:Macrophages / Q000276:immunology*; D013696:Temperature; D014357:Trypsin / Q000494:pharmacology
D008263:Macrophage Migration-Inhibitory Factors; D005643:Fucose; D014357:Trypsin; D005976:Glutaral