Skip to main content
SpringerLink
Log in

A Biotin–Avidin-Based Enzyme Immunoassay for βh-Endorphin

  • Published:
Pharmaceutical Research Aims and scope Submit manuscript

Abstract

An avidin–biotin enzyme-linked immunosorbent assay (ELISA) is described for βh-endorphin (βh-EP). Microtiter plates coated with commercially available antibodies were used together with βh-EP tracer derivatives that were biotinylated in positions 24, 28, and 29 via a C6 spacer arm. Nonspecific binding of biotinylated derivatives to the microtiter plates was blocked with a mixture of 1% casein and 10% ethanolamine in 0.1 M NaHCO3. A sequential saturation procedure using a high-affinity antiserum in combination with an avidin–alkaline phosphatase complex matched the sensitivity of reported radioimmunoassays (RIAs), with a detection limit of 0.5 fmol/assay. The intra- and interassay coefficients of variation were 5 and 12%, respectively. Results obtained by ELISA and RIA showed good correlations (r = 0.95). The β-EP concentration in extracted rat plasma after high-performance liquid chromatographic (HPLC) fractionation was determined by this method to be 1600 fmol/ml.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

REFERENCES

  1. J.-S. Hong, K. Yoshikawa, and R. W. Hendren. Methods Enzymol. 103:547–564 (1983).

    Google Scholar 

  2. J. Korpela. Med. Biol. 62:5–26 (1984).

    Google Scholar 

  3. E. A. Bayer and M. Wilchek. Methods Biochem. Anal. 26:1–45 (1980).

    Google Scholar 

  4. K. Wiedemann and H. Teschemacher. Pharm. Res. 3:142–149 (1986).

    Google Scholar 

  5. C. Cahill and H. Akil. Life Sci. 31:1871–1873 (1982).

    Google Scholar 

  6. G. Hochhaus, B. W. Gibson, and W. Sadee. J. Biol. Chem. 263:92–97 (1988).

    Google Scholar 

  7. H. Sato, Y. Sugiyama, Y. Sawada, T. Iga, and M. Hanano. Life Sci. 35:1051–1059 (1984).

    Google Scholar 

  8. J. Rossier, A. Bajon, T. M. Vargo, N. Ling, R. Guilleman, and F. Bloom. Life Sci. 21:847–852 (1977).

    Google Scholar 

  9. M.-T. Haeuptl, M. L. Aubert, J. Djiane, and J.-P. Kraehenbuhl. J. Biol. Chem. 258:305–314 (1983).

    Google Scholar 

  10. V. Hoellt, R. Przewlocki, and A. Herz. Naunyn-Schmiedeberg Arch. Pharmacol. 303:171–174 (1978).

    Google Scholar 

  11. J. K. Sarma, S. R. Hoffmann, and R. A. Houghten. Life Sci. 38:1723–173 (1986).

    Google Scholar 

  12. R. Guillemin, N. Ling, and T. Vargo. Biochem. Biophys. Res. Commun. 77:361–366 (1977).

    Google Scholar 

  13. J. Rossier, E. D. French, C. Rivier, N. Ling, R. Guillemin, and F. E. Bloom. Nature 170:618–620 (1977).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pharmacy, School of Pharmacy, University of California, San Francisco, California, 94143

    Guenther Hochhaus & Wolfgang Sadée

  2. Department of Pharmaceutics, College of Pharmacy, University of Florida, Gainesville, Florida, 32610

    Guenther Hochhaus

Authors
  1. Guenther Hochhaus
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wolfgang Sadée
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hochhaus, G., Sadée, W. A Biotin–Avidin-Based Enzyme Immunoassay for βh-Endorphin. Pharm Res 5, 232–235 (1988). https://doi.org/10.1023/A:1015993713471

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1015993713471

Search

Navigation