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Applications of Electrophoresis in studying New Antibiotics

Abstract

THE application of paper electrophoresis to the study of new antibiotics1 has been in use at these laboratories for some time. Our technique consists of applying spots of culture fluid, or a concentrate of it, to paper strips soaked in buffer, and subjecting the strips to electrophoresis by the technique of Kunkel and Tiselius2. Spots of chloromycetin are applied to the paper on each side of the spot of unknown antibiotic as a control for electro-osmosis. After migration, the position of the antibiotic is revealed by the microbiological technique as usual in paper chromatography of antibiotics3. Thus it is possible to determine whether unknown antibiotics in culture fluids are acidic, basic or amphoteric, and to obtain an approximate measure of their isoelectric point as a guide to subsequent purification procedures. The migration of an unknown antibiotic 179, relative to chloromycetin, to the cathode at pH 3 and pH 8, with lessened mobility at pH 8, indicates a basic substance with an isoelectric point greater than pH 8, as illustrated in the accompanying photograph.

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References

  1. Hoysoya, S., Soeda, M., Komatsu, N., Hara, N., Sonoda, Y., and Arai, R., J. Antibiot. Jap., 4, 314 (1951). Hoysoya, S., Soeda, M., Komatsu, N., Sonoda, Y., and Arai, R., J. Antibiot. Jap., 4, 317 (1951).

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  2. Kunkel, H. G., and Tiselius, A., J. Gen. Physiol., 35, 89 (1951).

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  3. Goodall, R. R., and Levi, A. A., Analyst, 72, 277 (1947). Winsten, W. A., and Eigen, E., J. Amer. Chem. Soc., 70, 3333 (1948). Peterson, D. H., and Reineke, L. M., J. Amer. Chem. Soc., 72, 3598 (1950).

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KING, N., DOERY, H. Applications of Electrophoresis in studying New Antibiotics. Nature 171, 878–879 (1953). https://doi.org/10.1038/171878b0

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