血清蛋白の「ポーラログラフ」的硏究(第1報) : I.「アルカリ」変性について

Abstract

We have studied the digest reaction of polarographic cancer tests (Brdička's), regarding its fundamental conditions of serum denaturation. Measuring the heights of protein double-wave obtained in cobaltous ammonium buffer solution, we called them H1 and H2 respectively. Since the H1 and H2 changed always correlatively and the change of H2 was greater than that of H1, we adopted the change of H2 as an indicator, and called the higher one "activated" and the lower ond "inactivated". Although the change of the height was almost negligible when serum was left at room temperature for a week, the height in 60℃ water-bath increased about 50% in 2 minutes and then decreased. In case of the alkaline denaturation (1/5N. KOH) at room temperature, the height was at its maximum in 5 minutes, about 2 times as the native, and then gradually decreased. After 3 hours, the height became lower than that of the control. In 60℃ waterbath this alkaline effect was so rapid that the height in 10 minutes corresponded to the in 24 hrs. Besides the temperature, the slope of this denaturation-time curve was found to be influenced regularly by KOH concentraton; the higher the concentration, the steeper the slope. The quantity of serum protein acted merely as a deciding factor of magnitude of this curve; but a protein-corrected test on a case of hypoproteinemia showed still lower wave height. Therefore, Brdička reaction is considered to be that of qualitative change after all. Furthermore, we found some interesting facts that the curve flattened and the peak (maximum activation) lagged in the pathologic sera, such as cancer or portal cirrhosis. From the above results, it may be said that the denaturation, especially alkaline one, might be a combination of two phases, i. e. phase of activation and that of inactivation, which will be characterized by temperature, agent and the nature of serum itself.