Development of a new radioimmunoassay for human osteocalcin: evidence for a midmolecule epitope.
Osteocalcin is a vitamin K-dependent bone-specific protein that can be found circulating in the serum. The circulating levels of osteocalcin have been shown to be an index of bone turnover. Existing radioimmunoassays for osteocalcin have been shown to be specific for C-terminal epitopes, a region that is identical in the human and bovine osteocalcin. There are, however, five amino acids different in the N-terminal region of the molecule. We describe here an immunoassay for a midmolecule epitope of osteocalcin using osteocalcin purified from human femoral head bone powder. Antibody specificity was determined using tryptic digests and a synthetic fragment of human osteocalcin. This assay has only a partial crossreactivity with bovine osteocalcin. This is the first report of an assay against a midmolecular epitope of osteocalcin involving a region in which the human and bovine osteocalcins differ. Osteocalcin levels determined by this assay have a significant correlation with both the total serum alkaline phosphatase and the serum skeletal alkaline phosphatase levels in normal adult human serum and, to a greater degree, in sera of patients with conditions associated with increased bone turnover (Paget's disease, hyperparathyroidism, and newborn sera). These correlations are greater than those previously reported for C-terminal assays, suggesting the possibility that different regions of the molecule may elicit different information concerning bone turnover.
Taylor
A K
AK
Jerry Pettis VA Hospital, Department of Biochemistry, CA, Loma Linda, CA 92357.
Linkhart
S G
SG
Mohan
S
S
Baylink
D J
DJ
D000328:Adult; D000469:Alkaline Phosphatase / Q000032:analysis; D000595:Amino Acid Sequence; D000918:Antibody Specificity; D001842:Bone and Bones / Q000032:analysis; D002135:Calcium-Binding Proteins / Q000097:blood* / Q000276:immunology / Q000302:isolation & purification; D000939:Epitopes / Q000032:analysis*; D006801:Humans; D008875:Middle Aged; D008969:Molecular Sequence Data; D015675:Osteocalcin; D011863:Radioimmunoassay
D002135:Calcium-Binding Proteins; D000939:Epitopes; D015675:Osteocalcin; D000469:Alkaline Phosphatase