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Development of an immunoassay for urinary galactosylhydroxylysine.

Abstract
Galactosylhydroxylysine (GHL) is released during bone resorption and has been shown to be elevated in subjects with metabolic bone loss. GHL is relatively specific for bone, it is not recycled or significantly metabolized during collagen turnover, and the levels are not influenced by diet. Previous measurements of GHL levels in urine have been performed using reverse-phase high performance liquid chromatography following pre-column derivatization. We produced polyclonal antibodies to GHL using GHL purified from sea sponges and developed an immunoassay that can recognize GHL in urine. The antibodies have minimal cross-reactivity with a physiological mixture of amino acids (< 1%), galactose (< 0.2%), lactose (< 0.3%), and glucosylgalactosylhydroxylysine (< 1%). This competitive immunoassay requires no dilution or pretreatment of the samples and provides a rapid and easy method for the evaluation of GHL in urine. Analysis of clinical samples from normal individuals, post-menopausal women, osteoporotic patients and individuals with Paget's disease show that the assay can discriminate between groups with differing levels of bone resorption as well as deoxypyridinoline (Dpd).
AuthorsS D Leigh, H S Ju, R Lundgard, G Y Daniloff, V Liu
JournalJournal of immunological methods (J Immunol Methods) Vol. 220 Issue 1-2 Pg. 169-78 (Nov 01 1998) ISSN: 0022-1759 [Print] Netherlands
PMID9839938 (Publication Type: Comparative Study, Journal Article)
Chemical References
  • Amino Acids
  • Biomarkers
  • Hydroxylysine
  • galactosylhydroxylysine
  • deoxypyridinoline
  • Collagen
Topics
  • Adult
  • Aged
  • Amino Acids (urine)
  • Animals
  • Biomarkers
  • Bone Resorption (diagnosis, urine)
  • Chromatography, High Pressure Liquid
  • Collagen (metabolism)
  • Female
  • Humans
  • Hydroxylysine (analogs & derivatives, immunology, isolation & purification, urine)
  • Immunoassay
  • Male
  • Middle Aged
  • Osteitis Deformans (urine)
  • Osteoporosis (metabolism)
  • Osteoporosis, Postmenopausal (urine)
  • Porifera (chemistry)
  • Postmenopause
  • Rabbits
  • Reference Values
  • Reproducibility of Results
  • Sensitivity and Specificity

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