Chemiluminescent Immunoassay for the Detection of Chloramphenicol Acetyltransferase and Human Growth Hormone Reporter Proteins

  • Corinne E. M. Olesen
  • John C. Voyta
  • Irena Bronstein
Part of the Methods in Molecular Biology™ book series (MIMB, volume 63)


Chemiluminescent substrates have been successfully used for highly sensitive quantitation of reporter enzyme activity (see  Chapters 5 and  6) and in enzyme-linked immunoassays for highly sensitive detection of many analytes (1, 2, 3, 4, 5, 6). CSPD chemiluminescent 1,2-dioxetane substrate (7) for alkaline phosphatase has been used with Sapphire-II enhancing reagent for enzyme-linked immunoassay detection of chloramphenicol acetyltransferase and human growth hormone reporter gene products. These chemiluminescent reporter gene assay systems provide nonradioactive, simple, sensitive detection methods, performed in microplate or tube luminometers.


Chloramphenicol Acetyltransferase Alkaline Phosphatase Conjugate Rubber Policeman Sensitive Detection Method Simultaneous Incubation 
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.


  1. 1.
    Bronstein, I., Voyta, J. C., Thorpe, G. H. G., Kricka, L. J., and Armstrong, G. (1989) Chemiluminescent assay of alkaline phosphatase applied in an ultrasensitive enzyme immunoassay of thyrotropin. Clin. Chem. 35, 1441–1446.PubMedGoogle Scholar
  2. 2.
    Thorpe, G. H. G., Bronstein, I., Kricka, L. J., Edwards, B., and Voyta, J. C. (1989) Chemiluminescent enzyme immunoassay of α-fetoprotein based on an adamantyl dioxetane phenyl phosphate substrate. Clin. Chem. 35, 2319–2321.PubMedGoogle Scholar
  3. 3.
    Bronstein, I., Voyta, J. C., Vant Erve, Y., and Kricka, L. J. (1991) Advances in ultrasensitive detection of proteins and nucleic acids with chemiluminescence novel derivatized 1,2-dioxetane enzyme substrates. Clin Chem. 37, 1526–1527.PubMedGoogle Scholar
  4. 4.
    Albrecht, S., Ehle, H., Schollberg, K., Bublitz, R., and Horn, A. (1991) Chemiluminescent enzyme immunoassay of human growth hormone based on adamantyl dioxetane phenyl phosphate substrate, in Bioluminescence and Chemiluminescence: Current Status (Stanley, P. and Kricka, L. J., eds.), John Wiley, Chichester, England, pp. 115–118.Google Scholar
  5. 5.
    Nishizono, I., Iida, S., Suzuki, N., Kawada, H., Murakami, H., Ashihara, Y., and Okada, M. (1991) Rapid and sensitive chemiluminescent enzyme immunoassay for measuring tumor markers. Clin. Chem. 37, 1639–1644.PubMedGoogle Scholar
  6. 6.
    Kricka, L. J. (1993) Ultrasensitive immunoassay techniques. Clin. Biochem. 26, 325–331.PubMedCrossRefGoogle Scholar
  7. 7.
    Bronstein, I., Juo, R. R., Voyta, J. C., and Edwards, B. (1991) Novel chemiluminescent adamantyl 1,2-dioxetane enzyme substrates, in Bioluminescence and Chemiluminescence: Current Status (Stanley, P. and Kricka, L. J., eds.), John Wiley, Chichester, England, pp. 73–82.Google Scholar
  8. 8.
    Gorman, C. M., Moffat, L. F., and Howard, B. H. (1982) Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol. Cell Biol. 2, 1044–1051.PubMedGoogle Scholar
  9. 9.
    Alam, J. and Cook, J. L. (1990) Reporter genes: Application to the study of mammalian gene transcription. Anal. Biochem. 188, 245–254.PubMedCrossRefGoogle Scholar
  10. 10.
    Young, S. L., Barbera, L., Kaynard, A. H., Haugland, R. P., Kang, H. C., Brinkley, M., and Melner, M. H. (1991) A nonradioactive assay for transfected chloramphenicol acetyltransferase activity using fluorescent substrates. Anal. Biochem. 197, 401–407.PubMedCrossRefGoogle Scholar
  11. 11.
    Burns, D. K. and Crowl, R. M. (1987) An immunological assay for chloramphenicol acetyltransferase. Anal. Biochem. 162, 399–404.PubMedCrossRefGoogle Scholar
  12. 12.
    Koch, W. and Burgelt, E. (1992) The CAT ELISA: An alternative nonradioactive approach for measuring gene expression. BMBiochemica. 9, 10a,b.Google Scholar
  13. 13.
    Boehringer Mannheim CAT ELISA Protocol, Cat No. 1363 727.Google Scholar
  14. 14.
    Hornbeck, P. (1994) Assays for antibody production, in Current Protocols in Immunology (Coligan, J. E., Kruisbeek, A. M., Margulies, D. H., Shevach, E. M., and Strober, W., eds.), Wiley, New York, p. 2.1.10.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1997

Authors and Affiliations

  • Corinne E. M. Olesen
    • 1
  • John C. Voyta
    • 1
  • Irena Bronstein
    • 1
  1. 1.TropixBedford

Personalised recommendations