Abstract
We have developed a sandwich-type enzyme immunoassay for human chorionic gonadotropin (hCG), in which antibody Fab’-β-D-galactosidase complex and an antibody-immobilized silicone rubber solid phase were used. Despite the fact that this assay system cross-reacted about 40% with human luteinizing hormone (LH) that contains an immunologically very similar subunit to that of hCG, it proved to be highly sensitive with hCG measurable at levels as low as 0.3 mlU per assay tube. Using 25 μl of serum sample or 100 μl of urine sample, hCG levels in serum (10–1000 mlU/ml) or in urine (3–300 mlU/ml) could be determined with the same degree of precision as in radioimmunoassay without sample interference with the assay. The coefficients of variation in within-run, and between-run were 9.2–13.3%, and 4.2–18.8%, respectively. Values obtained with enzyme immunoassay correlate well with those of radioimmunoassay (r = 0.961, slope = 1.129, y-intercept = 3.7 mlU/ml for 35 serum samples) and hemagglutination assay (r = 0.954, slope = 0.951, y-intercept = 1.8 mlU/ml for 88 urine samples).
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References
Midgley A.R. Radioimmunoassay; A method for human chorionic gonadotropin and human luteinizing hormone. Endocrinology 79: 10, 1966.
Tomoda Y., Hreschghyshyn M.M. Radioimmunoassay for human chorionic gonadotropin. Am. J. Obstet. Gynecol. 100: 118, 1968.
Van Weemen B.K., Schuurs A.H.W.N. Immunoassay using antigen-enzyme conjugates. FEBS Lett. 15: 232, 1971.
Van Weemen B.K., Schuurs A.H.W.N. Immunoassay using antibody-enzyme conjugates. FEBS Lett. 43: 215, 1974.
Yorde D.E., Sasse E.A., Wang T.Y., Hussa R.O., Garancis J.C. Competitive enzyme-linked immunoassay with use of soluble enzyme/antibody immune complexes for labelling. 1. Measurement of human choriogonadotropin. Clin. Chem. 22: 1372, 1976.
Kikutani M., Ishiguro M., Imamura S., Miura S. Enzyme immunoassay of human chorionic gonadotropin employing β-D-galactosidase as label. J. Clin. Endocrinol. Metab. 47: 980, 1978.
Van Hell H., Mattijsen R., Homan J.D.H. Studies on human chorionic gonadotropin: 1. Purification and some physico-chemical properties. Acta Endocrinol. 59: 89, 1968.
Kato K., Fukui H., Hamaguchi Y., Ishikawa E. Enzyme-linked immunoassay: Conjugation of the Fab fragment of rabbit IgG with β-D-galactosidase from E. Coli and its use for immunoassay. J. Immunol. 2116: 1554, 1976.
Kato K., Hamaguchi Y., Okawa S., Ishikawa E., Kobayashi K., Katunuma N. Use of rabbit antibody IgG-loaded silicone pieces for the sandwich enzymoimmunoassay of macromolecular antigens. J. Biochem. 81: 1557, 1977.
Bangham D.R., Grab B. The second International Standard for Chorionic Gonadotropin. Bull. WHO. 31: 111, 1964.
Kato K., Umeda Y., Suzuki F., Hayashi D., Kosaka A. Evaluation of a solid-phase enzyme immunoassay for insulin in human serum. Clin. Chem. 25: 1306, 1979.
Matuura S., Chen H.C., Hodgen G. Antibodies to the carboxyl-terminal fragment of human chorionic gonadotropin β-subunit: Characterization of antibody recognition sites using synthetic peptide analogues. Biochemistry 17: 575, 1978.
Vaitukaitis J.L., Braunstein G.D., Ross G.T. A radioimmunoassay which specifically measures human chorionic gonadotropin in the presence of human luteinizing hormone. Am. J. Obstet. Gynecol. 113: 751, 1972.
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Sekiya, T., Furuhashi, Y., Goto, S. et al. Sandwich-type enzyme immunoassay for human chorionic gonadotropin. J Endocrinol Invest 4, 275–279 (1981). https://doi.org/10.1007/BF03349444
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DOI: https://doi.org/10.1007/BF03349444