Combination of insulin chains on an anti-insulin antibody–Sepharose column


WHEN the mixture of products obtained by combining insulin A and B chains in solution (for a review, see ref. 1) was chromatographed on an anti-insulin antibody–Sepharose column2 no clear separation was achieved3. From the low biological activity (24.1%) of the material with the same antibody affinity as natural insulin (10.9% of the total bound products) it was concluded3 that resynthesised insulin with authentic structure constituted only a small portion of this fraction. Combination of insulin chains in solution has always involved a number of steps4,5 and has produced a mixture of products from which pure active insulin could be isolated by gel filtration and ion–exchange chromatography6. Linking the chains using the procedure described in this report, however, has given only one combination product that, according to various criteria, was identical with insulin and could be separated from unreacted A and B chains in one step.

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  1. 1

    Lübke, K., and Klostermeyer, H., Adv. Enzym. rel. Areas molec. Biol., 33, 445–525 (1970).

  2. 2

    Akanuma, Y., Kuzuya, T., Hayashi, M., Ide, T., and Kuzuya, N., Biochem. biophys. Res. Commun., 38, 947–953 (1970).

  3. 3

    Klotz, M., Diplomarbeit, Univ. Köln (1974).

  4. 4

    Brandenburg, D., Schermutzki, W., and Zahn, H., Hoppe-Seyler's Z. physiol. Chem., 354, 1521–1524 (1973).

  5. 5

    Geiger, R., and Obermeier, R., Biochem. biophys. Res. Commun., 55, 60–66 (1973).

  6. 6

    Rüegg, U. T., and Gattner, H.-G., Hoppe-Seyler's Z. physiol. Chem., 356, 1527–1533 (1975).

  7. 7

    Yagi, Y., Maier, P., and Pressman, D., Science, 147, 617–619 (1965).

  8. 8

    Gutte, B., thesis, Technische Hochschule Aachen (1967).

  9. 9

    Ouchterlony, O., Prog. Allergy, 5, 1–78 (1958).

  10. 10

    Levy, H. B., and Sober, H. A., Proc. Soc. exp. Biol. Med., 103, 250–252 (1960).

  11. 11

    Axen, R., Porath, J., and Ernback, S., Nature, 214, 1302–1304 (1967).

  12. 12

    Ellman, G. L., Archs Biochem. Biophys., 82, 70–77 (1959).

  13. 13

    Fruchter, R. G., and Crestfield, A. M., J. biol. Chem., 240, 3868–3874 (1965).

  14. 14

    Du, Y.-c., Zhang, Y.-s., Lu, Z.-x., and Tsou, C.-l., Scientia Sinica, 10, 84–104 (1961).

  15. 15

    Brown, H., Sanger, F., and Kitai, R., Biochem. J., 60, 556–565 (1955).

  16. 16

    Rodbell, M., J. biol. Chem., 239, 375–380 (1964).

  17. 17

    Gliemann, J., Diabetologia, 3, 382–388 (1967).

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