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Monoclonal antibodies specific for 18-hydroxycortisol and their use in an enzyme immunoassay for human urinary 18-hydroxycortisol for diagnosis of primary aldosteronism.

Abstract
We report the development of a monoclonal antibody-based enzyme immunoassay (EIA) specific for human urinary 18-hydroxycortisol, a biological marker of primary aldosteronism. Hybrid cell lines (hybridomas) were isolated after fusion between myeloma cells and spleen cells prepared from mice immunized with 18-hydroxycortisol conjugate. A competitive EIA suitable for the measurement of urinary 18-hydroxycortisol was developed using the mouse monoclonal antibody, KTM-41, which showed no practical cross-reaction with related endogenous steroids and synthetic steroids. This EIA meets all the requirements of routine clinical assay in terms of sensitivity (detection limit: 20 nmol/L), reproducibility (total CV: 8-15%), accuracy (recovery: 88-115%), simplicity and rapidity (< 3 h). Urinary 18-hydroxycortisol measured by the present assay was 153 +/- 119 nmol/L (mean +/- SD, range, 28-485) and 1787 +/- 1180 (range, 810-4264) in normal subjects (n = 20) and in patients with primary aldosteronism (n = 7), respectively. Clinical validation of the assay was confirmed by an appropriate decrease in urinary 18-hydroxycortisol level in patients with primary aldosteronism subsequent to adrenalectomy: 171 +/- 141 nmol/L (range, 41-466).
AuthorsH Kohno, S Sato, H Chiba, K Kobayashi, S Ikegawa, T Kurosawa, M Tohma
JournalClinical biochemistry (Clin Biochem) Vol. 27 Issue 4 Pg. 277-82 (Aug 1994) ISSN: 0009-9120 [Print] United States
PMID8001289 (Publication Type: Clinical Trial, Comparative Study, Journal Article, Randomized Controlled Trial)
Chemical References
  • Antibodies, Monoclonal
  • Biomarkers
  • 18-hydroxycortisol
  • Hydrocortisone
Topics
  • Animals
  • Antibodies, Monoclonal (immunology)
  • Antibody Specificity
  • Binding, Competitive
  • Biomarkers
  • Calibration
  • Chromatography, Affinity
  • Cross Reactions
  • Humans
  • Hybridomas
  • Hydrocortisone (analogs & derivatives, immunology, urine)
  • Hyperaldosteronism (diagnosis, urine)
  • Immunoenzyme Techniques
  • Male
  • Mice
  • Mice, Inbred BALB C

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