ネフロトキシン有効抗原の純化

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書誌事項

タイトル別名
  • The Purification of Nephrotoxic Serum Antigen
  • ネフロトキシン ユウコウ コウゲン ノ ジュンカ Starch zone electrophoresis オ モチイテ ノ ケントウ
  • -Studies with Starch Zone electrophoresis-
  • ― Starch Zone electrophoresisを用いての検討―

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説明

In the previous papers of our studies, it was demonstrated that the active principle of the soluble nephrotoxic serum antigen was contained in the polysaccharide moiety.<BR> However, considerable volume of nucleic acid and nitrogen were always found in the samples purifiedd by these methods.<BR> In this report, studies with zone electrophoresis were performed to remove the contaminative nucleic acid and to reduce the nitrogen level from the antigenic substance contained in the ultra supernatant of trypsin digested kidney homogenate.<BR> Starch block was divided into 4 sections (A: cathode side, B, C, D : anode sides), each of which was subjected to chemical analysis and immunopathological studies.<BR> The chemical composition of the fraction A showed nitrogen of about 5% and high hexose level. Faction B and C, on the contrary, showed high nitrogen and low hexose level. Nucleic acid was found only in fraction D.<BR> Antisera were prepared against each of these 4 sections through alum precipitation method, andd assayed. Only in rats injected with antisera to fraction A, the clinical and pathological changes of typical glomerulonephritis were induced. Therefore, it would appear that nephrotoxic serum antigenn was found only in fraction A.<BR> Fraction A was divided every 2 cm width and they were refered to fraction-3, fract. -5, fract. -7, fract. -9 and fract. -11, respectively.<BR> It was observed that in rats sacrificed 7 to 10 days after injection of antisera against fraction-3, clinical and pathological changes of kidneys were those of the severe glomerulonephritis. And further more, in rats sacrificed one month after injection of this antisera, histological changes of the kidneys were those of the typical contracted kidney.<BR> Slight activity was found in antisera to fractions-5 and-7.<BR> Chemical analysis of fraction-3 showed the highest hexose and low nitrogen level. The yield of fraction-3 was about 4 mg starting from 100 mg of ultrasupernatant.<BR> That is to say, the ultrasupernatant was purified by using of this zone electrophoresis method.<BR> And then, gel filtration of the antigenic substance on Sephadex columns G-50 and G-200 was performed<BR> Hexose and nitrogen were estimated. Fraction-3 showed a single peak at No. 4 fraction in Sephadex G-50, and at NO. 9 fraction in Sephadex G-200. These results would indicate that the effective antigenic substance was purified by using of the zone electrophoresis method.

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