合成基質を補助的に用いた補体C1の部分精製

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  • Purification of Complement Cl Monitored by a Nonhemolytic Activity Assay Using a Synthetic Substrate

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Various kinds of purification methods for the first complement component, Cl, have already been introduced, but most of them used gel-filtration or affinity chromatography after salting in and the Cl activity of the purified fractions were usually monitored by a hemolysis method. This paper describes an alternative method instead of a hemolytic method. For the activity assay, a synthetic substrate specific for Clr and Cls was used. In the purification steps, diisopropyl fluorophosphate or phenylmethylsulfonyl fluoride, inhibitors for serine proteases were added for gel-filtration. The former showed strong inhibitory effects on the natural activation of Cl during the purification steps, but the latter showed little effect. The biochemical characters of purified products were the same as those previously reported. Efficient utilization of proteases and synthetic substrates could provide good separation of inactive and active Cl components from the pooled plasma.

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