精製ツベルクリン(PPDs)とグルクロン酸ナトリウムとの結合-3-Sauton変法培地から得られたPPDsとグルクロン酸ナトリウム(NaG)との結合物(PPDs-G)の製法とその性状について
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In the previous papers, the authors reported that. PPDs from Sauton medium chemically combined with sodium glucuronate (NaG) under physiological conditions and that PPDs from modified Sauton medium would be preferable to that from Sauton medium, since physicochemical properties and tuberculin potencies of the former PPDs are invariable from. Lot to Lot.<BR>In order to prepare PPDs-G in homogeneous and reproducible state, we made PPDs from modified Sauton medium, and react them with NaG by simillar method as described in part I, and obtained PPDs-G. This paper presents the method of preparation of F'PDs-G and its physicochemical and biological properties.<BR>A mixture of PPDs and NaG (containing 5μC of 6-<SUP>14</SUP>C-NaG) in the ratio of 1 to 5 by weight in 0.2M phosphate buffer of pH 7.2 with a small amount of antiseptic were incubated at 38-40°C for 110-120 hours. The reaction mixture was then dialyzed against distilled water for 30 hours. Dialysate was condensed using Sephadex G 100 and applied to a column of Sephadex G 75. As seen in Fig. 1, elution curve of NaG was found to coincide with the first peak of PPDs, while by chromatography of a mixture of NaG and PPDs, the former could be completely separated from the latter. This suggests that PPDs combines with NaG. By calculation of NaG/PPDs in each fraction tube, it was found that the amount of NaG which was combined with PPDs would be at about 2-3% by weight of PPDs.<BR>Physicochemical properties of PPDs-G were as follows:<BR>(a) PPDs-G showed a little smaller value than PPDs in nitrogen content.<BR>(b) PPDs-G showed 2 bands in disc electrophoresis, using Tris-Glycin buffer of pH 8.6, one of which had a larger electrophoretic mobility than that of PPDs itself (Fig. 2). Isoelectric point of PPDs-G was estimated to be about 3.5-3. 7 by electric focusing in pH gradients (Fig. 3).<BR>(c) The reduction of basic amino acids content was observed in PPDs-G by amino acid analysis (Table 1).<BR>Biological properties of PPDs-G were as follows:<BR>(a) By tuberculin skin test, no significant difference was observed between the potencies of PPDs and PPDs-G.<BR>(b) The tuberculous guinea pigs were injected with either PPDs or PPDs-G. The number of dead animals in the group injected with PPDs-G seemed to be smaller than that in the group injected with PPDs (Table 2).<BR>(c) An attempt has been made by our coworkers to use PPDs-G as therapeutic agent against tuberculosis by combining them with chemotherapeutic agents. Preliminary study on the clinical effect is now under way.<BR>We further examined physicochemical properties of PPDs preparations from modified Sauton medium by measuring the contents, of nitrogen, sulfur, sugar, nucleic acids, or by ultracentrifugal analyses. As shown in Table 3, it was assumed that PPDs was not a pure substance as yet, but there was slight difference in physicochemical properties from Lot to Lot.
- 一般社団法人 日本結核病学会の論文
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- 結合物(PPDs-G)の製法ならびにその性状について (精製ツベルクリン(PPDs)とグルクロン酸ナトリウムとの結合-1,2-)
- 精製ツベルクリン(PPDs)とグルクロン酸ナトリウムとの結合-3-Sauton変法培地から得られたPPDsとグルクロン酸ナトリウム(NaG)との結合物(PPDs-G)の製法とその性状について