Chelated Terbium as a Label in Fluorescence Immunoassay

  • M. P. Bailey
  • B. F. Rocks
  • C. Riley


The use of fluorescent substances to covalently label biological molecules was described as early as 1941 by Coons and co-workers (Coons et al., 1941), who used anthracene isocyanate to label bacterial proteins. Later Coons and Kaplan (1950) introduced fluorescein isothiocyanate (FITC) as a more effective label. FITC has since become widely used as a fluorescent label in immunoassay. Its disadvantages are well recognised, however, and stem principally from the overlap of its emission spectrum with the endogenous fluorescence of plasma, and from its very small Stokes shift (only about 25 nm), which can lead to severe interference from scattered light.


Fluorescence Lifetime Fluorescence Immunoassay Excited State Population Homogeneous Assay Terbium Complex 
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  1. Bailey, M.P., Rocks, B.F. and Riley, C., 1986, Theoretical Considerations on the Lifetimes of Labels for Time-Resolved Fluoroimmunoassay, J. Pharm. Biomed. Anal., in press.Google Scholar
  2. Bailey, M.P., Rocks, B.F. and Riley, C., 1984, Terbium Chelate for Use as a Label in Fluoroimmunoassay, Analyst, 109: 1449–1450.PubMedCrossRefGoogle Scholar
  3. Bailey, M.P., Rocks, B.F. and Riley, C., 1985, Terbium Chelates for Fluorescence Immunoassay, Analyst, 110: 603–604.PubMedCrossRefGoogle Scholar
  4. Barela, T.D. and Sherry, D.A., 1976, A Simple, One-Step Fluorometric Method for Determination of Nanomolar Concentrations of Terbium, Anal. Biochem., 71: 351–357.PubMedCrossRefGoogle Scholar
  5. Butter, E., Kolowos, I. and Holzapfel, H., 1968, Ein Neues Reagenz zur Fluorometrischen Bestimmung von Terbium(III) und Dysprosium(III) in Wässriger Lösung, Talanta, 15: 901–911.CrossRefGoogle Scholar
  6. Coons, A.H., Creech, H.J. and Jones, R.N., 1941, Immunological Properties of an Antibody Containing a Fluorescent Group, Proc. Roy. Soc. Exp. Biol. (N.Y.), 47: 200–202.Google Scholar
  7. Coons, A.H. and Kaplan, M.H., 1950, Localization of Antigen in Tissue Cells.-II. Improvements in a Method for the Detection of Antigen by Means of Fluorescent Antibody, J. Exp. Med., 91: 1–13.PubMedCrossRefGoogle Scholar
  8. Haddad, P.R., 1977, The Application of Ternary Complexes to Spectrofluorometric Analysis, Talanta, 24: 1–13.PubMedCrossRefGoogle Scholar
  9. Hemmilä, I., Dakubu, S., Mukkala, V-M., Siitari, H. and Lövgren, T., 1984, Europium as a Label in Time-Resolved Immunofluorometric Assays, Anal. Biochem., 137: 335–343.PubMedCrossRefGoogle Scholar
  10. Hinshaw, J.C., Toner, J.L. and Reynolds, G.A., 1982, Eur. pat. appl. 82303380.8, Fluorescent Chelates and Labelled Specific Binding Reagents prepared therefrom.Google Scholar
  11. Lövgren, T., Hemmilä, I., Pettersson, K., Eskola, J.U. and Bertoft, E., 1984, Determination of Hormones by Time-Resolved Fluoroimmunoassay, Talanta, 31: 909–916.PubMedCrossRefGoogle Scholar
  12. Lyle, S.J. and Za’tar, N., 1983, A Comparative Study of Some Methods for the Spectrophotometric Determination of Terbium in Aqueous Solutions Containing Other Lanthanides and Yttrium, Anal. Chim. Acta, 153: 229–236.CrossRefGoogle Scholar
  13. Miller, J.N. and Thirkettle, C., 1975, The Fluorimetric Determination of Transferrin in Blood Serum, Biochem. Med., 13: 98–100.PubMedCrossRefGoogle Scholar
  14. Nargessi, R.D., Landon, J. and Smith, D.S., 1978, Non-separation Fluoroimmunoassay of Human Albumin in Biological Fluids, Clin. Chim. Acta., 89: 461–467.PubMedCrossRefGoogle Scholar
  15. Sidki, A.M. and Landon, J., 1985, Fluoroimmunoassay and Phosphoroimmunoassay, in “Alternative Immunoassays”, W.P. Collins, ed., J. Wiley, Chichester, 185–201.Google Scholar
  16. Sundberg, M.W., Meares, C.F., Goodwin, D.A. and Diamanti, C.I., 1974, Selective Binding of Metal Ions to Macromolecules Using Bifunctional Analogs of EDTA, J. Med. Chem., 17: 1304–1307.PubMedCrossRefGoogle Scholar
  17. Ullman, E.F., Schwarzberg, M. and Rubenstein, K.E., 1976, Fluorescent Excitation Transfer Immunoassay. A General Method for Determination of Antigens, J. Biol. Chem., 251: 4172–4178.PubMedGoogle Scholar
  18. Wieder, I., 1978, Background Rejection in Fluorescence Immunoassay, in: “Immunofluorescence and Related Staining Techniques. Proceedings of the VIth International Conference on Immunofluorescence and Related Staining Techniques.”, W. Knapp, K. Holubar and G. Wick, eds., Elsevier/North-Holland BiomEdical Press, Amsterdam and New York, 67–80, and references therein.Google Scholar
  19. Wilmott, N.J., Miller, J.N. and Tyson, J.F., 1984, Potential Use of a Terbium-Transferrin Complex as a Label in an Immunoassay for Gentamicin, Analyst, 109: 343–345.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • M. P. Bailey
    • 1
  • B. F. Rocks
    • 1
  • C. Riley
    • 1
  1. 1.Biochemistry DepartmentRoyal Sussex County HospitalSussexUK

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