Cited by 882Stress Responses and CortisolBiotin and Related StudiesAdrenal Hormones and Disorders

Synthesis of a cortisol–biotin conjugate and evaluation as a tracer in an immunoassay for salivary cortisol measurement

Dressendörfer, R.A., Kirschbaum, C., Rohde, W., Stahl, F., Strasburger, C.J.·1992·The Journal of Steroid Biochemistry and Molecular Biology

Abstract

Cortisol 3-(o-carboxymethyl)oxime (C3-CMO) and a commercially available biotin-hydrazide derivative were used to synthesize a C3-CMO-biotin conjugate. C3-CMO was converted into a N-hydroxysuccinimide ester derivative which in a second reaction step was allowed to interact with the hydrazide derivative of biotin. This simple-to-perform synthesis yielded a conjugate suitable for use as a tracer in immunoassays for cortisol measurement. Employing biotin as the primary probe in a competitive solid phase immunoassay allows for variable end point determination by means of commercially available labeled avidin or streptavidin derivatives. Streptavidin-Europium was used in conjunction with the DELFIA-system for time-resolved fluorometric end point measurement (TR-FIA) throughout the study. In addition, colorimetric end point determination (ELISA) using streptavidin-alkaline phosphatase as a secondary probe was established and evaluated. Both forms of this non-isotopic assay showed excellent correlation with a commercially available radioimmunoassay adapted for salivary cortisol measurement. The lower detection limit was 0.43 nM for a 50 microliters salivary sample. The intra-assay coefficient of variation was 6.7, 4.7 and 4.0% at cortisol concentrations of 2.2, 5.5 and 13.2 nM, respectively (n = 37), and the corresponding inter-assay coefficients of variation were 9.0, 8.6 and 7.1% (n = 50). The competitive immunoassay requires 1.5 h incubation time and shows robust and reproducible performance. The C3-CMO-biotin conjugate allows for sensitive and flexible end point determination of salivary cortisol levels in immunoassays.

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Crossref · 10.1016/0960-0760(92)90294-s

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  • Cortisol from salivettes was analyzed using a time-resolved fluorescence immunoassay with documented validity and reliability.

    “Cortisol from salivettes was analyzed using a time-resolved fluorescence immunoassay with documented validity and reliability (Dressendorfer et al., 1992).”
  • Assays were conducted in duplicate using a time-resolved immunoassay with fluorometric detection (DELFIA), with intra-assay coefficients of variation between 4.0% and 6.7% and inter-assay coefficients of variation from 7.1% to 9.0%.

    “Assays were conducted in duplicate using a time-resolved immunoassay with fluorometric detection (DELFIA) (Dressendorfer et al., 1992). Intra-assay coefficients of variation (CVs) were between 4.0% and 6.7%, and inter-assay CVs ranged from 7.1% to 9.0%.”

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Authors
Dressendörfer, R. A., Kirschbaum, C., Rohde, W., Stahl, F., & Strasburger, C. J.
Affiliations
  • LMU Klinikum (DE)
  • Universität Trier (DE)
  • Humboldt-Universität zu Berlin (DE)
Corresponding author
Strasburger, C.J.
Venue
The Journal of Steroid Biochemistry and Molecular Biology
Publisher
Elsevier BV
Published
1992-12
Volume
43
Issue
7
Pages
683-692
ISSN
0960-0760 (ISSN-L)
OpenAlex
W2036643530
Cited by
882
References
17
Citations by year
2026: 9 · 2025: 20 · 2024: 25 · 2023: 31 · 2022: 27 · 2021: 35 · 2020: 44 · 2019: 30 · 2018: 41 · 2017: 33 · 2016: 41 · 2015: 36 · 2014: 44 · 2013: 34 · 2012: 43
FWCI
2.84
Type
journal article
Language
en
Open access
closed
Locations
  • The Journal of Steroid Biochemistry and Molecular Biology (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Neuroscience › Behavioral Neuroscience › Stress Responses and Cortisol
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Biotin and Related Studies
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
Keywords
salivary cortisol, N-hydroxysuccinimide ester, competitive immunoassay, time-resolved fluoroimmunoassay, ELISA, steroid hormone measurement
MeSH terms
Biotin, Cross Reactions, Humans, Hydrocortisone, Immunoassay, Radioimmunoassay, Salivary Glands, Reproducibility of Results

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