Advertisement

Affinity Column Mediated Immunoenzymometric Assays

  • J. William Freytag

Abstract

Immunometric assays, by definition, are immunoassays configured in such a way that labeled-antibodies are utilized as the quantifying entity. This is in contrast to competitive radioimmunoassays or enzyme-immunoassays in which labeled-antigens are utilized in competition with sample antigen to bind to a limited number of antibody sites. The label applied to the antibody can take the form of a radioisotope, enzyme, fluorophore, luminescent tag, or almost any tag ultimately capable of generating a signal, even through some complex coupling mechanism. Two fundamentally different immunometric assay configurations have been described: the one-site immunometric assay and the two site (sandwich) immunometric assay.

Keywords

Affinity Column Unbind Fraction Immunometric Assay Crosslinking Reagent Control Pore Glass 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Avrameas, S. (1969). Coupling of enzymes to proteins with glutaraldehyde. Use of the conjugate for the detection of antigens and antibodies. Immunochemistry, 6: 43–52.PubMedCrossRefGoogle Scholar
  2. Avrameas, S. and T. Ternynck (1971). Peroxidase labeled antibody and Fab conjugates with enhanced intracellular penetration. Immunochemistry, 8: 1175–1179.PubMedCrossRefGoogle Scholar
  3. Engvall, E. and P. Perlmann (1971). Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry, 8: 871–874.PubMedCrossRefGoogle Scholar
  4. Freytag, J.W., J.C. Dickinson, and S.Y. Tseng (1984). A high sensitivity affinity-column-mediated immunometric assay, as exemplified by digoxin. Clinical Chemistry, 30: 417–420.PubMedGoogle Scholar
  5. Habermann, V.E. (1970). Ein neus prinzip zur quantitativen bestimmung hochmolekularer antigene. Z. klin. Chem. u. Klin. Biochem., 8: 51–55.Google Scholar
  6. Hashida, S., M. Imagawa, E. Ishikawa, and J.W. Freytag (1985). A simple method for the conjugation of affinity-purified Fab’ to horseradish peroxidase and β-galactosidase. Molecular Immunology (in press).Google Scholar
  7. Holbeck, S.L. and G.T. Nepon (1983). Enhanced detection of immunoglobulin binding by a modified ELISA. J. Immunol. Methods, 50: 47–52.CrossRefGoogle Scholar
  8. Imagawa, M., S. Hashida, E. Ishikawa, and J.W. Freytag (1984). Preparation of a monomeric 2,4-dinitrophenyl Fab’-β-galactosidase conjugate for immunoenzymometric assays. J. Biochem., 96: 1727–1735.PubMedGoogle Scholar
  9. Ishikawa, E., T. Kawai, and K. Miyai (1981). Enzyme Immunoassays. Igaku-Shoin, Tokyo. 280 pp.Google Scholar
  10. Ishikawa, E., M. Imagawa, S. Yoshitake, Y. Niitsu, I. Urushizaki, M. Inada, H. Imura, R. Kanazawa, S. Tachibana, N. Nakazawa, and H. Ogawa (1982). Major factors limiting sensitivity of sandwich enzyme immunoassay for ferritin, immunoglobulin E, and thyroid-stimulating hormone. Ann. Clin. Biochem., 19: 379–384.PubMedGoogle Scholar
  11. Ishikawa, E., M. Imagawa, S. Hashida, S. Yoshitake, Y. Hamaguchi, and T. Ueno (1983). Enzyme labeling of antibodies and their fragments for enzyme immunoassay immunohistochemical staining. J. Immunoassay, 4: 109–327.Google Scholar
  12. Leflar, C.C., J.W. Freytag, L.M. Powell, J.C. Strahan, J.J. Wadsley, C.A. Tyler, and W.K. Miller (1984). An automated affinity-column-mediated enzyme-linked immunometric assay for digoxin on the Du Pont aca® discrete clinical analyzer. Clinical Chemistry, 30: 1809–1811.PubMedGoogle Scholar
  13. Ling, M.C. (Feb. 18, 1974). United States Patent No. 3,867,517. Direct Radioimmunoassay for antigens and their antibodies.Google Scholar
  14. Maiolini, R., and R. Masseyeff (1975). A sandwich method of enzymoimmunoassay I. Application to rat and human alpha-fetoprotein. J. Immunol. Methods, 58: 293–300.Google Scholar
  15. Nomura, M., M. Imai, and K. Tasahashi (1983). Three-site radioimmunoassay with monoclonals for a sensitive determination of human alpha-fetoprotein. J. Immunol. Methods, 58; 293–300.PubMedCrossRefGoogle Scholar
  16. Pecht, I. (1982) Dynamic Aspects of Antibody Function. In The Antigen (M. Sela, Eds.). Academic Press, New York, NY. 33–42.Google Scholar
  17. Porstmann, B., T. Porstmann, R. Seifert, H. Meisel, and E. Nugel (1982). Comparison of direct and indirect two-site binding enzyme immunoassays. Clinica Chimica Acta, 122: 1–9.CrossRefGoogle Scholar
  18. Van Weeman, B.K. and A.H.W.N. Schuurs (1971). Immunoassays using antigen-enzyme conjugates. FEBS Letters, 15: 323–236.Google Scholar
  19. Weetall, H.H. (1973). Affinity Chromatography. Sep. and Purif. Meth., 2: 199–229.Google Scholar
  20. Yarmush, M.L. and C.K. Colton (1983). Affinity Chromatography. In Comprehensive Biotechnology. Pergamon Press.Google Scholar
  21. Yolken, R.H., F.J. Leister, L.S. Whitcomb, and M. Santosham (1983). Enzyme immunoassays for the detection of bacterial antigens utilizing biotin-labeled antibody and peroxidase biotin-avidin complex. J. Immunol. Methods, 56: 319–327.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • J. William Freytag
    • 1
  1. 1.Biomedical Products Department, Glasgow Research LaboratoryE. I. du Pont de Nemours & Co., Inc.WilmingtonUSA

Personalised recommendations