Methodology for the Study of Metabolism: Analyses of Proteins, Peptides, and Small Molecules

  • Jacob A. Canick


The ability to measure extremely low concentrations of biologically active molecules in fluid and tissue has enhanced the study of perinatal and neonatal metabolism as it has every other area of biology and medicine. Metabolic processes and biologic regulatory mechanisms are intimately connected, making the measurement of hormones and other biologic agents crucial to the study of metabolism. Even a cursory review of this textbook indicates the key role that such measurements have in a discussion of endocrine, paracrine, and/or autocrine effectors. To measure such effector molecules, both large and small, the technique of immunoassay is most commonly utilized, in which an antibody or group of antibodies can recognize and bind the molecule of interest with high specificity and great sensitivity.


Antibody Preparation Immunometric Assay Competitive Binding Assay Competitive Immunoassay Signal Antibody 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Yalow RS, Berson SA. Radiobiology. Assay of plasma insu lin in human subjects by immunological methods. Nature 1959; 184: 1648–1649.PubMedCrossRefGoogle Scholar
  2. 2.
    Yalow RS, Berson SA. Immunoassay of endogenous plasma insulin in man. J Clin Invest 1960; 39: 1157–1175.PubMedCrossRefGoogle Scholar
  3. 3.
    Berson SA, Yalow RS, Auerbach GD, et al. Immunoassay of bovine and human parathyroid hormone. Proc Natl Acad Sci USA 1963; 49: 613–617.PubMedCrossRefGoogle Scholar
  4. 4.
    Glick SM, Roth J, Yalow RS, et al. Immunoassay of human growth hormone in plasma. Nature 1963; 199: 784–787.PubMedCrossRefGoogle Scholar
  5. 5.
    Yalow RS, Glick SM, Roth J, et al. Radioimmunoassay of the human plasma ACTH. J Clin Endocrinol Metab 1964; 24: 1219–1225.PubMedCrossRefGoogle Scholar
  6. 6.
    Diamandis EP, Christopoulos TK, eds. Immunoassay. New York: Academic Press, 1996.Google Scholar
  7. 7.
    Wild D, ed. The immunoassay handbook. New York: Stockton Press, 1994.Google Scholar
  8. 8.
    Nakamura RM, Dasahara Y, Rechnitz GA, eds Immu nochemical assays and biosensor technology for the 1990s. Washington, DC: American Association for Microbiology, 1992.Google Scholar
  9. 9.
    Price CP, Newman DJ, eds. Principles and practice of immunoassay. New York: Stockton Press, 1991.Google Scholar
  10. 10.
    Collins WP, ed. Alternative immunoassays. New York: Wiley, 1985.Google Scholar
  11. 11.
    Kricka LJ, ed. Ligand-binder assays. New York: Marcel Dekker, 1985.Google Scholar
  12. 12.
    Albertson BD, Haseltine FP, eds. Non-radiometric assays. Technology and application in polypeptide and steroid hormone detection. New York: Alan R. Liss, 1988.Google Scholar
  13. 13.
    Jaffe BM, Behrman HR, eds. Methods of hormone radio-immunoassay. 2nd ed. New York: Academic Press, 1979.Google Scholar
  14. 14.
    Canick JA. Prenatal screening for Down syndrome using immunoassay. alpha-fetoprotein, unconjugated estriol, and human chorionic gonadotropin. J Clin Immunoassay 1990; 13: 30–33.Google Scholar
  15. 15.
    Porter P, Coley J, Gani M. Immunochemical criteria for successful matching of monoclonal antibodies to immu noassays of peptide hormones for assessment of pregnancy and ovulation. In: Albertson BD, Haseltine FP, eds. Non-radiometric assays. Technology and application in polypeptide and steroid hormone detection. New York: Alan R. Liss, 1988: 181–200.Google Scholar
  16. 16.
    Kohler G, Milstein C. Continuous culture of fused cells secreting antibody of predetermined specificity. Nature 1975; 256: 495–497.PubMedCrossRefGoogle Scholar
  17. 17.
    Kohler G, Milstein C. Derivation of specific antibody-producing tissue culture and tumour lines by cell fusion. Eur J Immunol 1976; 6: 511–519.PubMedCrossRefGoogle Scholar
  18. 18.
    Lieberman S, Erlanger BF, Beiser SM, et al. Aspects of steroid chemistry and metabolism. Steroid-protein conju gates: their chemical, immunochemical and endocrinologi- cal properties. Rec Progr Hormone Res 1959; 15: 165–200.Google Scholar
  19. 19.
    Erlanger BF. The preparation of antigenic hapten-carrier conjugates: a survey. Meth Enzymol 1980; 80: 85–104.CrossRefGoogle Scholar
  20. 20.
    Yalow RS. Radioimmunoassay of hormones. In: Wilson JD, Foster DW, eds. Williams’ textbook of endocrinology. 8th ed. Philadelphia: WB Saunders, 1992: 1635–1645.Google Scholar
  21. 21.
    Rodbard D, Lewald JE. Computer analysis of radioligand assay and radioimmunoassay data. Acta Endocrinol(Copenh) 1970;suppl 147: 7–103.Google Scholar
  22. 22.
    Healy MJR. Statistical analysis of radioimmunoassay data. Biochem J 1972; 130: 207–210.PubMedGoogle Scholar
  23. 23.
    Miles LEM, Hales CN. Labelled antibodies and immunological assay systems. Nature 1968; 219: 186–189.PubMedCrossRefGoogle Scholar
  24. 24.
    Miles LEM, Lipschitz DA, Bieber CP, et al. Measurement of serum ferritin by a 2-site immunoradiometric assay.Analyt Biochem 1974; 61: 209–224.CrossRefGoogle Scholar
  25. 25.
    Rodbard D, Feldman Y, Jaffe ML, et al. Kinetics of two-site immunoradiometric (“sandwich”) assays-II. Studies on the nature of “high-dose hook effect. ” Immunochemistry 1978; 15: 77–82.PubMedCrossRefGoogle Scholar
  26. 26.
    Schalch D, Parker M. A sensitive double antibody immu noassay for human growth hormone in plasma. Nature 1964; 203: 1141–1142.PubMedCrossRefGoogle Scholar
  27. 27.
    Dandliker WB, Feigen GA. Quantification of the antigenantibody reaction by the polarization of fluorescence. Biochem Biophys Res Commun 1961; 5: 299–304.PubMedCrossRefGoogle Scholar
  28. 28.
    Rubenstein KE, Schneider RS, Ullman EF. Homogeneous enzyme immunoassay. New immunochemical technique. Biochem Biophys Res Commun 1972; 47: 846–851.PubMedCrossRefGoogle Scholar
  29. 29.
    Schall RF Jr, Tenoso HJ. Alternatives to radioimmunoassay: labels and methods. Clin Chem 1981; 27: 1157–1164.PubMedGoogle Scholar
  30. 30.
    Howanitz JH. Immunoassay. Innovations in label technology. Arch Pathol Lab Med 1988; 112: 775–779.PubMedGoogle Scholar
  31. 31.
    Wisdom GB. Recent progress in the developmentof en zyme immunoassays. Ligand Rev 1981; 3: 44–49.Google Scholar
  32. 32.
    Hammilä I. Fluoroimmunoassays and immunofluorometric assays. Clin Chem 1985; 31: 359–370.Google Scholar
  33. 33.
    Diamandis EP. Immunoassays with time-resolved flourescence spectroscopy: principles and applications Clin Biochem 1988; 21: 139–150.PubMedGoogle Scholar
  34. 34.
    Weeks I, Woodhead JS. Chemiluminescence J Clin Immunoassay 1984; 7: 82–89.Google Scholar
  35. 35.
    Pandian MR, Morgan CH, Carlton E, et al. Modified immunoradiometric assay of parathyroid hormone-related peptide. Clinical application in the differential diagnosis of hypercalcemia. Clin Chem 1992; 38: 282–288.PubMedGoogle Scholar
  36. 36.
    Colton KW, Stevenson JC. Calcium metabolism. In: Wild D, ed. The immunoassay handbook. New York: Stockton Press, 1994: 362–365.Google Scholar
  37. 37.
    Jacobs DS, Kasten BL, Demott WR, et al. Laboratory test handbook. 2nd ed. Baltimore: Williams & Wilkins, 1990: 215.Google Scholar
  38. 38.
    Stone SJ, Henley R. The stability of blood samples for the measurement of the free beta subunit of chorionic gonadot- ropin. Prenat Diagn 1995; 15: 95–96.PubMedCrossRefGoogle Scholar
  39. 39.
    Baxter RC, Martin JL, Beniac VA. High molecular weight insulin-like growth factor binding protein complex. J Biol Chem 1989; 264: 11843–11848.PubMedGoogle Scholar
  40. 40.
    Rechler M. Insulin-like growth factor binding proteins.Vitam Horm 1993; 47: 1–114.PubMedCrossRefGoogle Scholar
  41. 41.
    Powell DR, Rosenfield RG, Baker BK, et al. Serum so-matomedin levels in adults with chronic renal failure: the importance of measuring insulin-like growth factor I (IGF- I) and IGF II in acid-chromatographed uriemic serum. J Clin Endocrinol Metab 1986; 63: 1186–1192.PubMedCrossRefGoogle Scholar
  42. 42.
    Underwood LE, Murphy MG. Radioimmunoassay of the somatomedins. In: Patrono C, ed. Radioimmunoassay in basic and clinical pharmacology (handbook of experimental pharmacology, vol. 82 ). Heidelberg: Springer-Verlag, 1987: 561–574.CrossRefGoogle Scholar
  43. 43.
    Lewitt MS, Denyer GS, Cooney GJ, et al. Insulin-like growth factor-binding protein-1 modulates blood glucose levels. Endocrinology 1991; 129: 2254–2256.PubMedCrossRefGoogle Scholar
  44. 44.
    Lee PDK, Powell D, Baker B, et al. Characterization of a direct, non-extraction immunoradiometric assay for free IGF-I. Endocrinology 1991; 129: 462A.Google Scholar
  45. 45.
    Smith RL. Effect of serum-separating gels on progesterone assays. Clin Chem 1985; 31: 1239.PubMedGoogle Scholar
  46. 46.
    Hilborn S, Krahn J. Effect of time of exposure of serum to gel-barrier tubes on results for progesterone and some other endocrine tests. Clin Chem 1987; 33: 203–204.PubMedGoogle Scholar
  47. 47.
    Hammond GL. Molecular properties of corticosteroid binding globulin and the sex-steroid binding proteins. Endocr Rev 1990; 11: 65–79.PubMedCrossRefGoogle Scholar
  48. 48.
    Cumming DC, Wall SR. Non-sex hormone binding globulin-bound testosterone as a marker for hyperandrogenism. J Clin Endocrinol Metab 1985; 61: 873–876.PubMedCrossRefGoogle Scholar
  49. 49.
    Ekins R. Measurement of free hormones in blood. Endocr Rev 1990; 11: 5–46.PubMedCrossRefGoogle Scholar
  50. 50.
    Pearce S, Dowsett M, Jeffcoate S. Three methods for esti- mating the fraction of testosterone and estradiol not bound to sex-hormone-binding globulin. Clin Chem 1989; 35: 632–635.PubMedGoogle Scholar
  51. 51.
    Fuqua JS, Sher ES, Migeon CJ, et al. Assay of plasma testosterone during the first six months of life: importance of chromatographic purification of steroids. Clin Chem 1995; 41: 146–1149.Google Scholar
  52. 52.
    Bartalena L. Recent achievements in studies on thyroid hormone-binding proteins. Endocr Rev 1990; 11: 47–64.PubMedCrossRefGoogle Scholar
  53. 53.
    Gruhn JG, Barsano CP, Kumar Y. The development of tests of thyroid function. Arch Pathol Lab Med 1987; 111: 84–100.PubMedGoogle Scholar
  54. 54.
    Dray F, Charbonnel B, Maclouf J.Radioimmunoassays for prostaglandins Fα, E1 and E2 in human plasma. Eur J Clin Invest 1975; 5: 311–318.PubMedGoogle Scholar
  55. 55.
    Kelly RW, Graham BJM, O’Sullivan MJ. Measurement of PGE2 as the methyl oxime by radioimmunoassay using a novel iodinated label. Prostaglandins Leukot Essent Fatty Acids 1989; 27: 187–191.CrossRefGoogle Scholar
  56. 56.
    Young DS, Bermes EW Jr. Specimen collection and pro cessing; sources of biological variation. In: Burtis CA, Ashwood CR, eds. Tietz textbook of clinical chemistry. 2nd ed. Philadelphia: WB Saunders, 1994: 58–101.Google Scholar
  57. 57.
    Westgard JO, Klee GG. Quality management. In: Burtis CA, Ashwood CR, eds. Tietz textbook of clinical chemistry. 2nd ed. Philadelphia: WB Saunders, 1994: 548–592.Google Scholar
  58. 58.
    Grammatopoulos D, Stirrat GM, Williams SA, et al. The biological activity of the corticotropin-releasing hormone receptor-adenylate cyclase complex in human myometrium is reduced at the end of pregnancy. J Clin Endocrinol Metab 1996; 81: 745–751.PubMedCrossRefGoogle Scholar
  59. 59.
    Crowther JR. ELISA: theory and practice. Methods in molecular biology, vol. 42. Totowa, NJ: Humana, 1995.Google Scholar
  60. 60.
    Ishikawa E. Development and clinical application of sensi tive enzyme immunoassay for macromolecular antigens—a review. Clin Biochem 1987; 20: 375–385.PubMedCrossRefGoogle Scholar
  61. 61.
    Moore GP. Genetically engineered antibodies. Clin Chem 1989; 35: 1849–1853.PubMedGoogle Scholar
  62. 62.
    Görög G, Gandolfi A, Paradisi G, et al. Use of bispecific hybrid antibodies for the development of a homogeneous enzyme immunoassay. J Immunol Methods 1989; 123: 131–140PubMedCrossRefGoogle Scholar
  63. 63.
    Ashihara Y, Nishizono I, Suzuki H, et al. Homogeneous enzyme immunoassay for macromolecular antigens using hybrid antibody. J Clin Lab Anal 1987; 1: 77–79.CrossRefGoogle Scholar
  64. 64.
    Davies C. Principles. In: Wild D, ed. The immunoassay handbook. New York: Stockton Press, 1994: 3–47.Google Scholar

Copyright information

© Springer Science+Business Media New York 1998

Authors and Affiliations

  • Jacob A. Canick

There are no affiliations available

Personalised recommendations