Effective purification of nonspecific cross-reacting antigens with phosphatidylinositol-specific phospholipase C.
Two molecular species of nonspecific cross-reacting antigens, NCA-90 and NCA-50 with mol. wts. of 90,000 and 50,000, respectively, were effectively extracted with phosphatidylinositol-specific phospholipase C (PI-PLC) from human lung tissues, followed by extraction with perchloric acid, immunoaffinity chromatography with anti-NCA adsorbent, and gel filtration on a TSK G3000SW column. The yields of NCA were about 2 times more than those obtained by the usual method without PI-PLC. Addition of 0.05 unit of PI-PLC to 1 g of lung tissue and incubation at 37 degrees C for 1 h with continuous shaking seem to be practically sufficient for NCA extraction. The immunochemical properties of the NCAs thus obtained were found to be identical to those of NCAs obtained by the ordinary method.
First Department of Biochemistry, School of Medicine, Fukuoka University, Japan.
M
M
Kuroki
M
M
Kuwahara
Y
Y
Tsuruta
M
M
Murakami
Y
Y
Matsuoka
D000941:Antigens / Q000302:isolation & purification*; D001344:Autopsy; D002272:Carcinoembryonic Antigen / Q000276:immunology / Q000302:isolation & purification; D002846:Chromatography, Affinity; D003429:Cross Reactions; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D007124:Immunoenzyme Techniques; D008168:Lung / Q000737:chemistry* / Q000276:immunology; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D054801:Phosphoinositide Phospholipase C; D010727:Phosphoric Diester Hydrolases
D000941:Antigens; D002272:Carcinoembryonic Antigen; D006023:Glycoproteins; D010727:Phosphoric Diester Hydrolases; D054801:Phosphoinositide Phospholipase C; D043265:Phosphatidylinositol Diacylglycerol-Lyase