Development of a monoclonal antibody to urinary degradation products from the C-terminal telopeptide alpha 1 chain of type I collagen. Application in an enzyme immunoassay and comparison to CrossLaps ELISA.
A monoclonal antibody MAbA7 was raised against a synthetic peptide having a sequence (EKAHDGGR) specific for a part of the C-telopeptide alpha 1 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 micrograms l-1 with a detection limit of 25 micrograms l-1. Within-run and total CVs were 5.5 and 8.0%, respectively. Analytical recovery averaged 96.6% +/- 5.3 (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.
Fledelius
C
C
Osteometer BioTech AS, Herlev, Denmark.
Kolding
I
I
Qvist
P
P
Bonde
M
M
Hassager
C
C
Reginster
J Y
JY
Hejgaard
J
J
Frøokiaer
H
H
Christiansen
C
C
D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D000918:Antibody Specificity; D001862:Bone Resorption / Q000276:immunology / Q000652:urine; D003094:Collagen / Q000737:chemistry / Q000276:immunology* / Q000652:urine*; D024042:Collagen Type I; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D008875:Middle Aged; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology* / Q000652:urine*; D010455:Peptides / Q000737:chemistry / Q000276:immunology* / Q000652:urine*; D017698:Postmenopause; D017697:Premenopause
D000911:Antibodies, Monoclonal; D024042:Collagen Type I; D010446:Peptide Fragments; D010455:Peptides; C052929:collagen type I trimeric cross-linked peptide; D003094:Collagen