Juntendo Medical Journal

Studies on the Phage Protein. (IV) Antibody Production with P22 Coat Protein. Michiko HORIKOSHI and Kiyoaki KAMIJO Tissue Culture Lab. ndo University, School of Medicine. Immunological activities of the P22-protein was described previously. Comparative study was performed on the immune response of the P22-protein, original P22 phage and host protein (LT 2 protein). Experiments were carried cut with rabbits. Results obtained were as follows ; 1. In a neutralizing antibody response, it must be emphasized that the P22-protein seemed to be equivalent to the original P22 phage. 2. In process of the antibody production, however, some difference was seen between anti-P22-protein- and anti-P22 -sera, suggesting the existence of some different factor in P22-protein from the original phage. It may be the degenerative product in degradation process or the changes in tertiary or quatery structure of the coat protein. 3. In animals injected with LT 2 protein (host protein), the antibody produced was quite different from those against P22 or P22-protein. The host protein should be different nature of the phage protein in immunological activity. 4. For the neutralizing antibody production, the necessary and enough doses of antigens were 5 x 10¹¹-1.15 X 10¹² for P22 phage and 2.0-2.63 mg for P22 protein, respectively. These doses should be given in 4 or 5 times.

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Studies on the phage protein (V). Yields of the three fractions through DEAF-cellulose column chromatography and the immunological activities of them. Michiko Horikoshi and Kiyoaki Kamijo Tissue Culture Laboratory, Juntendo University, School of Medicine The P22 phage protein was fractionated through DEAE-cellulose column and the three fractions, F 1, F 2 and F 3 were obtained successfully as described in previous paper. Yields of the three fractions were estimated under various conditions and the further investigations were carried out with them on immunological and physicochemical properties. Results were concluded as follows. 1. Average yields of the fractions were 27.5%, 0.9% and 2.6% for F1, F2 and F3, respectively. Maximal yields were obtained with 1.0g of DEAE-cellulose and 0.2ml/min of the flow-rate, i. e., 59%, 24% and 4.4%, respectively. 2. The yields decreased with the more amount of DEAE-cellulose and increased with the slower flow rate, generally. 3. In blocking tests, F3 was exceedingly effective and was followed by F1 and F2. 4. The precipitin titer was highest in anti-F1 serum but the relative value per mg of antigen injected was greatest in anti-F3 serum and was followed by anti-F2 and -F1 sera. F3 was more effective than P22-protein, in this respect, whereas the latter exceeded slightly the former in the blocking power. 5. Neutralizing antibody was produced equally but in less degree than previous report with any of the fractions. This phenomenon should be retested because the protein amount injected was far less in cases of F2 and F3 than of F1. From the results of the blocking test, it is strongly sugested that the neutralizing antibody, also, will be produced exceedingly in case of F3. 6. In immunoelectrophoresis, the relative migration of the 3 fractions coincided well to those in polyacrylamide gel electrophoresis and the other components than three fractions could not be found.

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