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Fecal Glucocorticoid Analysis: Non-invasive Adrenal Monitoring in Equids
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Highly Responsive Bioassay for Quantification of Glucocorticoids.

Mathias Flensted Poulsen1, Martin Overgaard2,3, Christian Brix Folsted Andersen1

  • 1Department of Biomedicine, Aarhus University, Wilhelm Meyers Allé 3, 8000 Aarhus C, Denmark.

Analytical Chemistry
|January 26, 2024
PubMed
Summary

A new cell-based assay, HEK293F-GRE, accurately measures total and bioavailable cortisol in serum. This versatile tool simplifies bioavailable cortisol measurement, aiding personalized glucocorticoid therapy.

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Area of Science:

  • Endocrinology
  • Biochemistry
  • Assay Development

Background:

  • Current methods for measuring total cortisol, like immunoassays and LC-MS/MS, are established.
  • Measuring bioavailable cortisol is currently complex, unreliable, and inconvenient for patients.

Purpose of the Study:

  • To develop and validate a novel cell-based assay for measuring both total and bioavailable cortisol in serum.
  • To provide a more convenient and reliable method for assessing cortisol status.

Main Methods:

  • Development of a human embryonic kidney cell line reporter assay (HEK293F-GRE).
  • Validation of the assay for stability, accuracy, precision, repeatability, reproducibility, and specificity.
  • Testing the assay's performance in measuring total and bioavailable cortisol in human serum samples.

Main Results:

  • The HEK293F-GRE assay demonstrated linearity (R² = 0.98) and accuracy (0.8-1.2) across dilutions.
  • Assay precision was validated down to 3-6 nM dexamethasone (CV < 20%).
  • Total cortisol measurements correlated well with immunoassay and LC-MS/MS; bioavailable cortisol estimations aligned with literature values.

Conclusions:

  • The HEK293F-GRE assay is a promising complementary method for clinical cortisol assessment.
  • Direct quantification of bioavailable cortisol facilitates personalized glucocorticoid dosing regimens.