Precipitation method for separating and quantifying bone and liver alkaline phosphatase isoenzymes.
We evaluated a method for quantifying bone isoenzyme of alkaline phosphatase (ALP) which utilizes wheat-germ lectin to precipitate this fraction. In precision studies, CVs ranged from 3.2 to 11.4% (within-day) and from 3.7 to 11.5% (day-to-day). The assay procedure was linear to 1100 U/L and was easily adapted to automated kinetic measurement. Comparison of the precipitation method with an affinity electrophoretic method, which utilizes cellulose acetate as a support, demonstrated a satisfactory coefficient of correlation (r = 0.886). The reference range was determined in sera from 188 healthy adult subjects. The distribution of bone ALP values was also studied in 73 healthy children and in 30 healthy adolescents. To evaluate the clinical applicability of the method, the bone isoenzyme was determined in samples from several groups of subjects (pregnant women, patients with hepatobiliary diseases, patients with hepatocellular carcinoma without skeletal involvement, and patients with bone, liver or lymph node metastases). We found the method suitable for routine determination of bone alkaline phosphatase and for the screening of bone metastases. Because of its technical simplicity and satisfactory analytical performance, it can be used instead of the heat-inactivation procedure.
Burlina
A
A
Centro di Ricerca Biomedica della Regione Veneto, Università di Padova, Italy.
Plebani
M
M
Secchiero
S
S
Zaninotto
M
M
Sciacovelli
L
L
D000293:Adolescent; D000328:Adult; D000368:Aged; D000469:Alkaline Phosphatase / Q000302:isolation & purification* / Q000378:metabolism; D001842:Bone and Bones / Q000201:enzymology*; D011232:Chemical Precipitation; D002648:Child; D002675:Child, Preschool; D004796:Clinical Enzyme Tests / Q000379:methods*; D004588:Electrophoresis, Cellulose Acetate; D005069:Evaluation Studies as Topic; D005260:Female; D006801:Humans; D007223:Infant; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D008297:Male; D008875:Middle Aged; D011247:Pregnancy; D012016:Reference Values; D014909:Wheat Germ Agglutinins
D007527:Isoenzymes; D014909:Wheat Germ Agglutinins; D000469:Alkaline Phosphatase