Effective purification of nonspecific cross-reacting antigens with phosphatidylinositol-specific phospholipase C.

M Kuroki, M Kuwahara, Y Tsuruta, M Murakami, Y Matsuoka
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引用次数: 3

Abstract

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.

用磷脂酰肌醇特异性磷脂酶C有效纯化非特异性交叉反应抗原。
两种非特异性交叉反应抗原,NCA-90和NCA-50与molwts。分别用磷脂酰肌醇特异性磷脂酶C (PI-PLC)从人肺组织中有效提取9万个和5万个,然后用高氯酸提取,抗nca吸附剂免疫亲和层析,在TSK G3000SW柱上凝胶过滤。NCA的产率是不加PI-PLC的常规方法的2倍左右。在1 g肺组织中加入0.05单位的PI-PLC,在37℃下连续摇动培养1小时,似乎足以提取NCA。由此获得的NCAs的免疫化学性质与用普通方法获得的NCAs相同。
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