Development of a sensitive, direct luminescent enzyme immunoassay for plasma estradiol-17β

A. Roda, S. Girotti, A. L. Piacentini, S. Preti, S. Lodi

Research output: Contribution to journalArticlepeer-review

Abstract

A rapid, sensitive, precise, chemiluminescent enzyme immunoassay for estradiol-17β has been developed and validated. Antibodies were produced in rabbits using estradiol-17β-6-(O-carboxymethyl)oxime coupled to bovine serum albumin, purified and immobilized on polystyrene beads (6.4 mm diameter). The same derivative was used to prepare the enzymatic tracer by coupling with horseradish peroxidase. The assay, direct on the serum sample, featured a 4-h binding step at 4°C followed by the chemiluminescent detection using luminol/H2O2. The detection limit was 0.15 pg/tube and the assay was carried out on 20-100 μl of sample, allowing measurement of estradiol-17β in plasma concentrations from 1.5 to 500 pg/ml. The method fulfills all the standard requisites of precision and accuracy and the results agree well with a radioimmunoassay procedure on extracted serum.

Original languageEnglish
Pages (from-to)267-273
Number of pages7
JournalAnalytical Biochemistry
Volume156
Issue number2
DOIs
Publication statusPublished - Aug 1 1986

Keywords

  • estradiol-17β
  • luminescent enzyme immunoassay
  • radioimmunoassay

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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