腎トリプシン消化超遠心上清 (Nephrotoxin 有効抗原) 作製條件の検討

概要

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It is previously reported that the ultrasupernatant of trypsin digested rabbit (rat, dog) kidney cortex homogenate contained nephrotoxic complete antigen in a soluble form, and chemically consisted of the nearly constant levels of nitrogen, hexose, hexosamine, phosphorus etc. The original method for extracting this active antigenic substance from kidney cortex homogenate required 100 mg(50 mg) trypsin and three hours incubation in 37℃ for the digestion of 12g (6g) kidney homogenate. But the minimum requirements for trypsin quantity and incubation time were not yet discussed. Experiments were tried in this paper to determine the minimum conditions for extracting active antigenic substance from 6g dog kidney homogenate. When trypsin≧30 mg and incubation time of three hours were given, the chemical cmpositions of the ultrasupernatants were nearly identical with that of samples extracted by the original method. On the other hand, when trypsin≧20 mg and incubation time of three hours were given, chemical properties of the ultrasupernatants were lower than normal range of samples by the original method. Incubation should be continued at least for one hour, even if trypsin≧30 mg was given. The curve of the trypsic digesting process measured using the Kunitz method supported the results above mentioned. When non-digested kidney homogenate was centrifuged and then its sediment was digested with trypsin, more efficient curve was shown.
1966-07-30