Improved purification of human bone sialoprotein and development of a homologous radioimmunoassay

Bone sialoprotein (BSP) is a phosphorylated skeletal glycoprotein. Here we describe a new procedure for the purification of BSP involving wide-pore reversed-phase HPLC, and the development of a homologous RIA for human BSP. The immunoassay showed linearity between 3 and 120 micrograms/L, a lower det...

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Bibliographic Details
Published in:Clinical chemistry (Baltimore, Md.) Md.), 1997-11, Vol.43 (11), p.2076-2082
Main Authors: Karmatschek, Markus, Maier, Ina, Seibel, Markus J, Woitge, Henning W, Ziegler, Reinhard, Armbruster, Franz P
Format: Article
Language:English
Subjects:
Adult
Aged
Aged, 80 and over
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Chromatography, High Pressure Liquid
Drug Stability
Femur - chemistry
Fundamental and applied biological sciences. Psychology
Glycoproteins
Humans
Hyperthyroidism - blood
Integrin-Binding Sialoprotein
Investigative techniques, diagnostic techniques (general aspects)
Iodine Radioisotopes
Kidney Failure, Chronic - blood
Liver Cirrhosis, Alcoholic - blood
Medical sciences
Middle Aged
Miscellaneous. Technology
Osteitis Deformans - blood
Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques
Proteins
Radioimmunoassay - methods
Sialoglycoproteins - blood
Sialoglycoproteins - isolation & purification
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Summary:Bone sialoprotein (BSP) is a phosphorylated skeletal glycoprotein. Here we describe a new procedure for the purification of BSP involving wide-pore reversed-phase HPLC, and the development of a homologous RIA for human BSP. The immunoassay showed linearity between 3 and 120 micrograms/L, a lower detection limit of 0.7 micrograms/L, and a mean recovery rate of 99.4%. Intraassay variation was 7.0% (mean = 10.9 micrograms/L) and 6.1% (mean = 38.8 micrograms/L), and interassay variation was 9.2% (mean = 11.1 micrograms/L) and 9.4% (mean = 39.0 micrograms/L). No cross-reactivity was detected with osteocalcin, osteonectin, or bone alkaline phosphatase. Preliminary clinical evaluation in healthy controls (n = 90) showed a mean serum BSP concentration on 12.1 +/- 5.0 micrograms/L (+/- SD). BSP was significantly increased in patients with Paget disease of bone, primary and secondary hyperparathyroidism, and also in subjects with renal failure without skeletal involvement. Impairment of hepatic function did not affect serum BSP concentrations.
ISSN:0009-9147
1530-8561
DOI:10.1093/clinchem/43.11.2076