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Original

Development of a monoclonal antibody to urinary degradation products from the C-terminal telopeptide α1 chain of type I collagen. Application in an enzyme immunoassay and comparison to CrossLaps™ ELISA

, , , , , , , & show all
Pages 73-83 | Received 09 May 1996, Accepted 25 Nov 1996, Published online: 29 Mar 2011
 

Abstract

A monoclonal antibody MAbA7 was raised against a synthetic peptide having a sequence (EKAHDGGR) specific for a part of the C-telopeptide α1 chain of type I collagen. MAbA7 was labelled with horseradish peroxide and used in a competitive one-step enzyme-linked immunosorbent assay (ELISA) for measurement of urinary type I collagen degradation products. The assay was technically evaluated and preliminary clinical data are presented. The measuring range was 200–7000 µg 1−1 with a detection limit of 25 µg 1−1. Within-run and total CVs were 5.5 and 8.0%, respectively. Analytical recovery averaged 96.6%±M5.3 (mean±1SD). Values obtained in the ELISA were highly correlated (r=0.93) to values obtained by a commercially available assay (CrossLaps™ ELISA) known to measure urinary degradation products derived from the C-telopeptide of type I collagen reflecting the rate of bone resorption. Investigation of the urinary fragments responsible for the immunological response in the two assays revealed, however, that they are not identical. Values obtained in urine samples from postmenopausal women (n=108) and patients with Paget's disease (n=6) increased 43% (p<0.01) and 28-fold (p<0.001), respectively, when compared to a premenopausal level (n=50). A decrease in the urinary concentrations of 67% (p<0.01) was seen after 6 months in urine samples from postmenopausal women (n = 13) receiving hormone replacement therapy (HRT) compared to a group receiving placebo (n=9). Likewise, the urinary concentrations decreased 88% (p<0.001) in early postmenopausal women receiving bisphosphonate therapy (n = 11) for a period of 9 months compared to a group receiving placebo (n = 12). These results suggest that the monoclonal antibody and the new assay may be useful for further investigations of the physiological and clinical importance of type I collagen degradation.

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