Summary
Solid phase biochemical systems have many technical advantages. The lecture reviews a number of ways in which solid phase immunological methods may be used in diabetes research. Solid phase antibodies are very convenient for use in radioimmunoassay. The immunoradiometric assay involves the isolation of antibodies onto solid phase antigen, the iodination of the antibodies whilst combined to the solid phase antigen, the elution of the iodinated antibodies and finally their use as a reagent for the measurement of antigen. The two site assay utilises solid phase antibodies to extract and concentrate antigen from solutions such as plasma. The extracted antigen may then be assayed whilst combined with solid phase antigen if a second site on the antigen is available for reaction with I125 — labelled antibody. High specific activity, purified labelled antibodies may also be used for antigen localization, antibody estimation and labelling cell membranes. Cell membrane purification using solid phase antibodies is considered as a further development of this methodology.
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Addison, G.M., Hales, C.N., Miles, L.E.M.: Principles and modification of radioimmunological assays of protein hormones. Horm. Metab. Res. Suppl. 3 Eds. Federlin, K., Hales, C.N., Kracht, J., pp. 22–25, Stuttgart: George Thieme Verlag 1971.
— —: The immunoradiometric assay. In: Radioimmunoassay methods. Eds. Kirkham, K.E., Hunter, W.M. pp. 447–461. London: Churchill Livingstone 1971a.
— —: Discussion paper in: Radioimmunoassay methods. Eds. Kirkham, K.E., Hunter, W.M., pp. 481–487. London: Churchill Livingstone 1971b.
— —: Two site assay of human growth hormone. Horm. Metab. Res.3, 59–60 (1971c).
- Beamish, M.R., Hales, C.N., Hodgkins, M., Jacobs, A., Llewellyn, P.: An immunoradiometric assay for ferritin in serum: normal subjects and patients with iron deficiency and iron overload. J. clin. Path, in Press (1972).
Berson, S.A., Yalow, R.S., Bauman, S.A., Rothschild, M.A., Newerly, K.: Insulin-I131 metabolism in human subjects: demonstration of insulin binding globulin in the circulation of insulin treated subjects. J. clin. Invest.35, 170–190 (1956).
Catt, K.J.: Radioimmunoassay with antibody — coated discs and tubes. Acta endocr. Suppl.142, 222–243 (1969).
Cuatreeasas, P., Anfinsen, C.B.: Purification of biologically active proteins and peptides by affinity chromatography. In: Methods in Enzymology21. Eds. Colowick, S.P., Kaplan, N.O. New York: Academic Press (In press) 1971.
Greenwood, F.C., Hunter, W.M., Glover, J.S.: The preparation of I131 labelled human growth hormone of high specific radioactivity. Biochem. J.89, 114–123 (1963).
Gurvich, A.E., Kuzovleva, O.B., Tumanova, A.E.: Production of protein-cellulose complexes (immunosorbents) in the form of suspensions able to bind great amounts of antibodies. Biokhimiya26, 803–809 (1961).
Hales, C.N., Randle, P.J.: Immunoassay of insulin with insulin — antibody precipitate. Biochem. J.88, 137–146.
- unpublished observations (1972).
McFarlane, A.S.: Efficient trace — labelling of proteins with iodine. Nature (Lond.)182, 53.
Mettler, C.C.: History of Medicine, pp. 467–469. Philadelphia: The Blaekiston Company 1947.
Miles. L.E.M., Hales, C.N.: Labelled antibodies and immunologieal assay systems. Nature (Lond.)219, 186–189 (1968a).
— —: The preparation and properties of purified125I-labelled antibodies to insulin. Biochem J.108, 611–618 (1968b).
— —: Immunoradiometric assay procedures: new developments. In: In vitro procedures with radioisotopes in medicine. Proceedings of the Symposium on In vitro Procedures with Radioisotopes in Clinical Medicine and Research organised by the International Atomic Energy Agency in co-operation with the World Health Organisation, pp. 483–489. Vienna: International Atomic Energy Agency 1970.
—: Isolation of specific high affinity antibodies from nonspecific antisera. J. clin. Endocr.33, 399–408 (1971).
Moloney, P.J., Coval, M.: Antigenicity of insulin. Diabetes induced by specific antibodies. Biochem. J.59, 179–185 (1955).
O'Riordan, J.L.H., Addison, G.M., Woodhead, J.S., Keutman H.T., Potts J.T.: Localisation of antigenic sites in parathyroid hormone. In: Endocrinology 1971 proceedings of a symposium held in London July 1971. Ed. Taylor, S.F. London: Heinemann Medical Books Ltd., in press (1972).
Schlichtkrull, J.: Paper read at the 7th Annual Meeting of the European Association for the Study of Diabetes, Southampton (1971).
Shapiro, E., Strauss, L.: Oskar Minkowski's registration of left atrial pressure curves in mitral valve disease. Amer. J. Cardiol.14, 107–111 (1964).
Steiner, D.F., Clark, J., Nolan, C., Rubenstein, A.H., Margoliash, E., Aten, B., Oyer, P.: Proinsulin and the biosynthesis of insulin. Recent Progr. Hormone Res.25, 207–272 (1969).
Stewart, J.M., Young, J.D.: Solid phase peptide synthesis. San Francisco: W.H. Freeman and Company 1969.
Yalow, R.S., Berson, S.A.: Assay of plasma insulin in human subjects by immunologieal methods. Nature (Lond.)184, 1648 (1959).
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The studies reported here were supported by the British Diabetic Association, the British Insulin Manufacturers and the Medical Research Council.
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Hales, C.N. Immunological techniques in diabetes research. Diabetologia 8, 229–235 (1972). https://doi.org/10.1007/BF01225565
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DOI: https://doi.org/10.1007/BF01225565