Studies on the Mechanism of Antibody Formation (9)

An adequate amount of typhoid-paratyphoid vaccine (TPV) or bovine liver lipid which had been emulsified in bovine serum was injected intravenously to rabbits of two groups two times at an interval of 4 days. After the second sensitization, it was found that a new specific substance had appeared in the circulating blood of each rabbit, and that it had accelerated the endogenous respiration of mouse liver cells remarkably under aerobic conditions.An effective serum was used as a starting material and examined first for stability against heat, acid, and alkali and dialyzability agains semi-dialytic cellophane membrane. Then, the extraction and purification were carried out by the column-chromatographic method. In this method, the deproteinized serum filtrate was absorbed on Amberlite IR 120H+ resin with 100-200 meshes. After that, the effective substance was extracted as a single substance through the column-chromatographic fractionation by effusion of 0.1N-HCl solution. Finally, it was identified as uric acid by paper chromatography, continuous ultraviolet and infrared absorption spectra, element dialysis, and measurement of melting point. On the other hand, uric acid was compared with some metabolic intermediates of ATP, with the exception of uric acid itself, as substrates for the oxidative respiration of mouse liver homogenate or mitochondria.