Quantification of skeletal alkaline phosphatase in osteoporotic serum by wheat germ agglutinin precipitation, heat inactivation, and a two-site immunoradiometric assay.

Three methods for quantifying skeletal alkaline phosphatase (ALP; EC 3.1.3.1) activity/immunoactivity in serum--heat inactivation, wheat germ agglutinin (WGA) precipitation, and an immunoradiometric assay--were tested for recovery and specificity and applied to 81 sera collected from 14 postmenopausal osteoporotic subjects. The heat-inactivation and WGA precipitation assays showed relative recoveries of 91-100% and 16-32%, respectively, for skeletal ALP with complete specificity (no cross-reactivity with hepatic or intestinal ALP); the IRMA showed a relative recovery of 86-100% and 11-14% cross-reactivity with hepatic ALP. There was a closer correlation between the heat-inactivation assay and IRMA (r = 0.833) than between the WGA precipitation assay and IRMA (r = 0.673) or between the heat-inactivation and WGA-precipitation assays (r = 0.568). The WGA precipitation assay failed to detect skeletal ALP in three serum samples that contained significant amounts as determined by the heat-inactivation assay and the IRMA.

[1]  J. Griffiths,et al.  Separation and identification of alkaline phosphatase isoenzymes and isoforms in serum of healthy persons by isoelectric focusing. , 1987, Clinical chemistry.

[2]  T. Kuwana,et al.  Sugar-chain analysis of alkaline phosphatase by lectin affinity chromatography in serum from a patient with osteosarcoma. , 1989, Clinical chemistry.

[3]  J. Wergedal,et al.  Reference standards for quantification of skeletal alkaline phosphatase activity in serum by heat inactivation and lectin precipitation. , 1993, Clinical chemistry.

[4]  J. Wergedal,et al.  Skeletal alkaline phosphatase specific activity is an index of the osteoblastic phenotype in subpopulations of the human osteosarcoma cell line SaOS-2. , 1991, Metabolism: clinical and experimental.

[5]  C. W. Small.  A study of the heat inactivation of serum alkaline phosphatase by an automated technique. , 1969, Clinica chimica acta; international journal of clinical chemistry.

[6]  W. Behr,et al.  Quantification of bone alkaline phosphatase in serum by precipitation with wheat-germ lectin: a simplified method and its clinical plausibility. , 1986, Clinical chemistry.

[7]  M. Peake,et al.  Quantitative method for determining serum alkaline phosphatase isoenzyme activity I. Guanidine hydrochloride: new reagent for selectively inhibiting major serum isoenzymes of alkaline phosphatase. , 1986, Journal of clinical pathology.

[8]  A. Y. Foo,et al.  Two new methods for separating and quantifying bone and liver alkaline phosphatase isoenzymes in plasma. , 1984, Clinical chemistry.

[9]  D. Baylink,et al.  Skeletal alkaline phosphatase activity as a bone formation index in vitro. , 1986, Metabolism: clinical and experimental.

[10]  M. Ferguson,et al.  Cell-surface anchoring of proteins via glycosyl-phosphatidylinositol structures. , 1988, Annual review of biochemistry.

[11]  G. Valenzuela,et al.  Time-dependent changes in bone, placental, intestinal, and hepatic alkaline phosphatase activities in serum during human pregnancy. , 1987, Clinical chemistry.

[12]  H. N. Fernley,et al.  Studies on alkaline phosphatase. Inhibition of human-placental phosphoryl phosphatase by L-phenylalanine. , 1972, European journal of biochemistry.

[13]  G. Wesolowski,et al.  Characterization of a human osteosarcoma cell line (Saos-2) with osteoblastic properties. , 1987, Cancer research.

[14]  W. Fishman,et al.  Automated differential isoenzyme analysis. II. The fractionation of serum alkaline phosphatases into "liver", "intestinal" and "other" components. , 1971, Enzymologia.

[15]  H. van Belle.  Alkaline phosphatase. I. Kinetics and inhibition by levamisole of purified isoenzymes from humans. , 1976, Clinical chemistry.

[16]  I. Koyama,et al.  Differences between human liver- and bone-type alkaline phosphatases. , 1989, Clinica chimica acta; international journal of clinical chemistry.

[17]  J. Reginster,et al.  New radioimmunoassay for determination of bone specific iso-enzymes of serum alkaline phosphatases activity (Ostase°): validation and clinical perspective , 1993 .

[18]  W. Fishman.  Alkaline phosphatase isozymes: recent progress. , 1990, Clinical biochemistry.

[19]  J. S. Clubb,et al.  HEAT INACTIVATION IN THE STUDY OF HUMAN ALKALINE PHOSPHATASES. , 1965, Annals of internal medicine.

[20]  J. O'brien,et al.  Alkaline phosphatase isoenzymes of liver and bone origin are incompletely resolved by wheat-germ-lectin affinity chromatography. , 1989, Clinical chemistry.

[21]  J. Katzmann,et al.  Isolation and preliminary characterization of a monoclonal antibody that interacts preferentially with the liver isoenzyme of human alkaline phosphatase. , 1985, Clinical chemistry.

[22]  C. Alvarez Moreno,et al.  Comparison of three bone alkaline phosphatase quantification methods in patients with increased alkaline phosphatase activities. , 1992, Clinical chemistry.

[23]  B. Riggs,et al.  Clinical evaluation of high-performance affinity chromatography for the separation of bone and liver alkaline phosphatase isoenzymes. , 1991, Clinica chimica acta; international journal of clinical chemistry.

[24]  K. Fedde.  Human osteosarcoma cells spontaneously release matrix-vesicle-like structures with the capacity to mineralize. , 1992, Bone and mineral.

[25]  I. Ramasamy,et al.  Wheat germ lectin affinity electrophoresis of serum alkaline phosphatase with commercially available agarose gels. , 1993, Clinical chemistry.

[26]  J. Farley,et al.  Quantitation of skeletal alkaline phosphatase isoenzyme activity in canine serum , 1992, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[27]  J. Farley,et al.  Alkaline phosphatase activity from human osteosarcoma cell line SaOS-2: an isoenzyme standard for quantifying skeletal alkaline phosphatase activity in serum. , 1989, Clinical chemistry.

[28]  M. Kaplan.  Alkaline phosphatase. , 1972, Gastroenterology.

[29]  M. Dambacher,et al.  Bone remodeling and calcium metabolism: a correlated histomorphometric, calcium kinetic, and biochemical study in patients with osteoporosis and Paget's Disease. , 1977, Metabolism: clinical and experimental.

[30]  H. Belle.  Alkaline phosphatase. I. Kinetics and inhibition by levamisole of purified isoenzymes from humans. , 1976 .

[31]  P. Crofton.  Wheat-germ lectin affinity electrophoresis for alkaline phosphatase isoforms in children: age-dependent reference ranges and changes in liver and bone disease. , 1992, Clinical chemistry.

[32]  R. Wolfert,et al.  The preparation of monoclonal antibodies which react preferentially with human bone alkaline phosphatase and not liver alkaline phosphatase. , 1990, Clinica chimica acta; international journal of clinical chemistry.

[33]  M. Peake,et al.  Quantitative method for determining serum alkaline phosphatase isoenzyme activity: estimation of intestinal component. , 1988, Journal of clinical pathology.

[34]  W. E. Schreiber,et al.  Alkaline phosphatase isoenzymes resolved by electrophoresis on lectin-containing agarose gel. , 1986, Clinical chemistry.

[35]  L. Whitby,et al.  A simplified heat-inactivation method for investigating alkaline phosphatase isoenzymes in serum. , 1975, Clinica chimica acta; international journal of clinical chemistry.

[36]  H. Anderson.  VESICLES ASSOCIATED WITH CALCIFICATION IN THE MATRIX OF EPIPHYSEAL CARTILAGE , 1969, The Journal of cell biology.

[37]  D. Moss.  Perspectives in alkaline phosphatase research. , 1992, Clinical chemistry.

[38]  A. Saltiel,et al.  Structural and functional roles of glycosyl-phosphatidylinositol in membranes. , 1988, Science.

[39]  E. Schoenau,et al.  Liquid-chromatographic determination of isoenzymes of alkaline phosphatase in serum and tissue homogenates. , 1986, Clinical chemistry.

[40]  H. Sussman,et al.  Characterization of a human osteoblastic osteosarcoma cell line (SAOS‐2) with high bone alkaline phosphatase activity , 1987, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.

[41]  W. Fishman,et al.  L-Homoarginine. An organ-specific, uncompetitive inhibitor of human liver and bone alkaline phosphohydrolases. , 1972, The Journal of biological chemistry.

[42]  G. Lum,et al.  Clinical assessment of the electrophoretic separation of alkaline phosphatase isoenzymes. , 1983, American journal of clinical pathology.

[43]  C. Chesnut,et al.  Improved method for quantitative determination in serum of alkaline phosphatase of skeletal origin. , 1981, Clinical chemistry.

[44]  P. Mayne,et al.  Evaluation of two new methods for routine measurement of alkaline phosphatase isoenzymes. , 1992, Journal of clinical pathology.

[45]  K. Mann,et al.  Concurrent assays of circulating bone Gla-protein and bone alkaline phosphatase: effects of sex, age, and metabolic bone disease. , 1988, The Journal of clinical endocrinology and metabolism.

[46]  J. Fennelly,et al.  The Value of Differential Alkaline Phosphatase Thermostability in Clinical Diagnosis , 1969 .