9.補体成分の分離と力価測定法(第3回補体シンポジウム I 補体成分の基礎的研究)

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Methods for separation, purification with DEAE and CM cellulose columns, and measurement of nine components of guinea pig complement were recently established by R.A. Nelson, J.Jensen, I.Gigli, and N.Tamura. The nine components of complement in our laboratory were separated from guinea pig serum with a little modified system. Each of the separated components had more than 1,000 C'H50 in activity, but the contamination of other components was less than 30 C'H50, if it were. The inactive protein contained in the separated components was extremely low except C'2 and C'3e. Using these components, measurement system was built up, i.e. C'1:EA4, C'2, EDTA-C; C'4:EAI, C'2, EDTA-C'; C'2:EA14, DETA-C';, C'3c: EA142, C'3 befad(IH), or EA142, HuE(IA); C'3b: EA1423c, C'3 efad; C'3e: EA1423c, C'3 bfad; C'3f: EA1423c, C'3bead; C'3a: EA1423 cbef, C'3d; C'3d; EA1423 cbef, C'3a. Especially in titration of C'3 group, the same effective molecule of the seprated components was used in the assay system and this assay system could provide reproducible maximum endtiter of each component. With the assay system, nine components in Geon zone electrophoresis and Cohn's ethanol fractions were measured. Most of them were found in β-globulin region and C'3d, 3b, 3e were slightly in the faster fraction and other C'3 components were in the slower fraction. Dose response curve and site number was determined in each step. Provided site number of EA14 was about 70 per cell, site number of SA1423 cbef would have been about 50 per cell. Dose response curve was linar in each step except in terminal step. These results proved that titration of C'3c, b, e, f and a could be performed with intermediate cell and titration with intermediate cell would be superior to other assay system in analysis of kinetics and inhibition. But EA1423cb cell is extremely unstable and titration of C'3e was performed with C'3e site generation. In this case titer of C'3e was reproducible and maximal. On titration of 3c with IA and IH, there was a remarkable difference between them. Studied on the same sites in number higher dilution of C'2 would be more sensitive in IA, and these resuts suggested that high concentration of C'2 would bring out immune hemolysis effectively. Under these experiment C'2 fraction enhanced the immune hemolysis, especially after EA1423 cbefa cell stage and multiple nature of C'3 activity was discussed.
日本アレルギー学会の論文
1966-11-30

9.補体成分の分離と力価測定法(第3回補体シンポジウム I 補体成分の基礎的研究)

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