Immunoradiometric method and electrophoretic system compared for quantifying bone alkaline phosphatase in serum

1995 ◽  
Vol 41 (6) ◽  
pp. 853-857 ◽  
Author(s):  
V O Van Hoof ◽  
M Martin ◽  
P Blockx ◽  
A Prove ◽  
A Van Oosterom ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Malignant Disease ◽  
Screening Method ◽  
Bone Alkaline Phosphatase ◽  
Cross Reactivity ◽  
Osteoblastic Activity ◽  
Alp Activity ◽  
Agarose Electrophoresis ◽  
End Stage ◽  
Rule Out

Abstract Agarose electrophoresis (Isopal, Beckman) and an immunoradiometric assay (IRMA) involving specific monoclonal antibodies (Ostase, Hybritech), two methods for the quantification of serum bone alkaline phosphatase (ALP, EC 3.1.3.1), a marker of osteoblastic activity, were compared in 293 patients: 79 with end-stage renal failure treated with hemodialysis and 214 with malignant disease. Overall correlation between the two methods was good (r = 0.92), except (a) for low values of bone ALP and (b) in some samples with high total liver ALP activity--both due to considerable cross-reactivity of the anti-bone ALP antibodies of the Ostase kit with liver ALP. This interference was not constant and was not evenly distributed across all concentrations of bone ALP. Low bone ALP determined with the IRMA (< or = 5 micrograms/L) was confirmed by electrophoresis (< or = 21 U/L), but bone ALP activity determined by electrophoresis to be low (< or = 21 U/L) was not correlated with the IRMA results. After standardizing our results by computing z-values for bone ALP, delta z (= zOstase - zIsopal) was significantly correlated with liver ALP activity (r = 0.73, P < 0.0001). We conclude that the IRMA for quantifying bone ALP is acceptable as a screening method. However, when high values for bone ALP are found with the Ostase method, confirmation by electrophoresis remains mandatory to rule out cross-reactivity with high amounts of liver ALP. For detecting low bone ALP activities, electrophoresis remains the method of choice.

scholarly journals Method for determination of bone alkaline phosphatase activity: analytical performance and clinical usefulness in patients with metabolic and malignant bone diseases

1996 ◽  
Vol 42 (2) ◽  
pp. 210-217 ◽  
Author(s):  
W Withold ◽  
U Schulte ◽  
H Reinauer
Keyword(s):  
Alkaline Phosphatase ◽  
Bone Alkaline Phosphatase ◽  
Cross Reactivity ◽  
Bone Diseases ◽  
Characteristic Analysis ◽  
Diagnostic Efficacy ◽  
Alp Activity ◽  
Osteoblast Activity ◽  
Score Analysis

Abstract We report the performance characteristics of an assay for determination of bone alkaline phosphatase (ALP) activity after immunoadsorption in microplate wells. Between-run imprecision was between 7.1% and 11.2%. The detection limit was 1.0 U/L. Comparisons with an immunoradiometric test for determination of bone ALP mass concentrations yielded the following regression equation: y = 3.11 + 1.33x with y, the bone ALP activity concentration (U/L) (and x, the bone ALP mass concentration microgram/L) (r +=0.974, n = 103). Using sera from patients with liver diseases and sera from patients with secondary hyperparathyroidism yielded a cross-reactivity of 20% for circulating liver ALP (and its membrane-bound isoform). In patients receiving renal transplants, Z-score analysis revealed that after transplantation the increase in bone ALP activity is more pronounced than total ALP activity. In tumor patients, receiver-operating characteristic analysis revealed that bone ALP activity shows the same diagnostic efficacy as total ALP activity in the detection of bone metastases (as assessed by bone scintigraphy). In multiple myeloma patients, suppressed osteoblast activity was well detectable by bone ALP activity determination.

Alkaline Phosphatase Isoenzyme Patterns in Malignant Disease

1992 ◽  
Vol 38 (12) ◽  
pp. 2546-2551 ◽  
Author(s):  
V O Van Hoof ◽  
A T Van Oosterom ◽  
L G Lepoutre ◽  
M E De Broe
Keyword(s):  
Alkaline Phosphatase ◽  
Liver Disease ◽  
Liver Metastasis ◽  
Bone Metastasis ◽  
Positive Predictive Value ◽  
Bone Disease ◽  
Predictive Value ◽  
Malignant Disease ◽  
Alp Activity ◽  
Isoenzyme Patterns

Abstract Early treatment of patients with malignant disease and liver or bone metastasis may increase their survival time. We have used the activity patterns of liver and bone isoenzymes of alkaline phosphatase (ALP), separated by agarose gel electrophoresis, to detect early metastasis. We studied ALP isoenzyme patterns in a background population of 101 patients with no evidence of any disease that might influence this pattern; a healthy reference population (n = 330); and the following three groups of patients: 143 with malignant disease, 47 with nonmalignant liver disease, and 22 with nonmalignant bone disease. Cutoff and predictive values of liver ALP, high-molecular-mass (high-M(r)) ALP, and bone ALP were established for detecting liver and bone metastasis. The positive predictive value of liver and high-M(r) ALP was higher than that of total ALP in detecting liver metastasis, but liver and high-M(r) ALP did not enable us to differentiate between malignant and nonmalignant liver disease. Total ALP activity was of slightly more value than liver and high-M(r) ALP in enabling us to rule out liver metastasis. From bone ALP activity we could not distinguish between nonmalignant bone disease and bone metastasis. The negative predictive value of bone ALP in the diagnosis of bone metastasis was low, but its positive predictive value was high and superior to that of total ALP.

scholarly journals Immunoadsorption assay for bone alkaline phosphatase compared with wheat germ agglutinin precipitation assay in serum from (pre)pubertal girls

1996 ◽  
Vol 42 (12) ◽  
pp. 1970-1974 ◽  
Author(s):  
A A Bouman ◽  
C M de Ridder ◽  
J H Nijhof ◽  
J C Netelenbos ◽  
H A Delemarre-vd Waal
Keyword(s):  
Alkaline Phosphatase ◽  
Correlation Coefficient ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Bone Alkaline Phosphatase ◽  
Cross Reactivity ◽  
Reference Ranges ◽  
Correlation Studies ◽  
Deming Regression ◽  
The Mean

Abstract The performance characteristics of two bone alkaline phosphatase (ALP; EC 3.1.3.1) assays, a wheat germ agglutinin (WGA) precipitation assay and a new immunoadsorption assay (IAA), were compared. The within- and between-run imprecision of the IAA (3.6-4.2% and 3.6-7.7%) was comparable with that of the WGA assay. The mean cross-reactivity with liver ALP appeared to be 4% in the WGA assay and 11% in the IAA. The reference ranges in a group of 155 healthy Caucasian (pre)pubertal schoolgirls were: 149-401 U/L (total ALP, 30 degrees C), 105-349 U/L (bone ALP, 30 degrees C, WGA assay), and 58-205 U/L (bone ALP, 25 degrees C, IAA). Comparison of the WGA assay (x) with the IAA (y) demonstrated a correlation coefficient of 0.95 [Deming regression equation: y = (0.56 +/- 0.01)x + (2.0 +/- 1.5); Sy[symbol: see text]x = 5.3 U/L]. Correlation studies of the WGA assay and the IAA results with total ALP demonstrated r = 0.98 and 0.96, respectively.

Bone alkaline phosphatase on the IDS-iSYS automated analyser; cross-reactivity with intestinal ALP

2019 ◽  
Vol 57 (8) ◽  
pp. e186-e188 ◽  
Author(s):  
Jennifer J. Brady ◽  
Desmond McGoldrick ◽  
Kate O’Callaghan ◽  
Fiona McNamara ◽  
Keith J. Mulready ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Bone Alkaline Phosphatase ◽  
Cross Reactivity

Biochemical Changes following Parathyroidectomy

1993 ◽  
Vol 16 (10) ◽  
pp. 700-703 ◽  
Author(s):  
P.K.G. Chandran ◽  
T.J. Ulahannan ◽  
M. Skiles
Keyword(s):  
Alkaline Phosphatase ◽  
Serum Calcium ◽  
Serum Alkaline Phosphatase ◽  
Inorganic Phosphorus ◽  
Biochemical Changes ◽  
Postoperative Hypocalcemia ◽  
Subtotal Parathyroidectomy ◽  
Osteoblastic Activity ◽  
End Stage ◽  
Phosphorus Levels

Biochemical changes that had appeared after subtotal parathyroidectomy (PTx) in 26 patients with end-stage renal failure were observed. The volume of excised parathyroid glands was also measured. Serum calcium and inorganic phosphorus levels fell after PTx; only to rise in due course. Serum alkaline phosphatase levels rose after PTx, reaching a peak by the 14th postoperative day. These elevated levels returned to normal range at about three months after PTx. Strong correlation was noted among the degree of postoperative hypocalcemia, and increase in serum alkaline phosphatase, but not between absolute pre or postoperative alkaline phosphatase levels and changes in serum calcium or phosphorus concentrations. Nevertheless, significant correlation was seen between pre-PTx levels of alkaline phosphatase and its short-lived postoperative rise, indicating hastened osteoblastic activity.

scholarly journals Mass versus Activity: Validation of an Immunometric Assay for Bone Alkaline Phosphatase in Serum

1995 ◽  
Vol 32 (4) ◽  
pp. 405-412 ◽  
Author(s):  
C P Price ◽  
C A Mitchell ◽  
J Moriarty ◽  
M Gray ◽  
K Noonan
Keyword(s):  
Renal Failure ◽  
Alkaline Phosphatase ◽  
Reference Range ◽  
Paget’S Disease ◽  
Bone Alkaline Phosphatase ◽  
Paget's Disease ◽  
Cross Reactivity ◽  
Age Group ◽  
Immunometric Assay ◽  
Activity Measurements

A detailed investigation of the performance of an immunometric assay for the quantitation of bone alkaline phosphatase (ALP) in serum showed that precision of this assay was similar to that of lectin precipitation and electrophoretic methods. Relationships for mass and activity measurements of the bone isoform using samples from children and patients with Paget's disease were similar for lectin and electrophoretic methods. Purified liver enzyme showed 100% cross reactivity in the immunometric assay. However, comparison of the slopes between bone ALP mass and total ALP activity using only samples with predominantly liver or bone isoforms showed that the cross reactivity of the liver isoform in serum was 18·3%. Experiments in which increasing amounts of a sample containing 90% of liver isoform adding to a serum sample from a patient with Paget's disease showed a cross reactivity of 16·5%. The reference range for bone ALP mass was 7–28 μg/L for men ( n = 77) and 5–20 μg/L for women ( n = 110) in the age group 20–50 years. In women over 50 years bone ALP was 28% higher. Increased bone ALP mass was also demonstrated in patients with Paget's disease ( n = 59), liver disease ( n = 95), chronic renal failure ( n = 41) and hyperthyroidism ( n = 17).

Isolation and preliminary characterization of a monoclonal antibody that interacts preferentially with the liver isoenzyme of human alkaline phosphatase.

1985 ◽  
Vol 31 (3) ◽  
pp. 381-385 ◽  
Author(s):  
G M Lawson ◽  
J A Katzmann ◽  
T K Kimlinger ◽  
J F O'Brien
Keyword(s):  
Monoclonal Antibody ◽  
Alkaline Phosphatase ◽  
Human Liver ◽  
Clinical Laboratory ◽  
Bone Alkaline Phosphatase ◽  
Cross Reactivity ◽  
Immunological Methods ◽  
Enzymic Assay ◽  
Murine Monoclonal Antibodies

Abstract We have prepared murine monoclonal antibodies against isolated human bone alkaline phosphatase (ALP, EC 3.1.3.1). Hybridoma supernates were separately screened for reactivity against both human liver and bone ALP. Although most antibody-positive hybrids showed similar reactivity against both isoenzymes, one hybridoma produced an antibody that interacted preferentially with liver ALP. This antibody was purified and used to establish an immunoassay to differentiate liver ALP from bone ALP. When equal activities of the two isoenzymes (as determined by a conventional enzymic assay) were measured by the immunoassay, a fivefold greater response was obtained with liver than with bone ALP. The immunoassay can be used to measure the proportions of the bone and liver isoenzymes in mixtures of them. Cross reactivity with human placental and intestinal ALP is less than 3% relative to liver ALP. These findings support the feasibility of developing immunological methods to differentiate these isoenzymes in the clinical laboratory.

Age and sex distribution of alkaline phosphatase isoenzymes by agarose electrophoresis

1990 ◽  
Vol 36 (6) ◽  
pp. 875-878 ◽  
Author(s):  
V O Van Hoof ◽  
M F Hoylaerts ◽  
H Geryl ◽  
M Van Mullem ◽  
L G Lepoutre ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Age Groups ◽  
Soluble Form ◽  
Alp Activity ◽  
Normal Individuals ◽  
Bone Fraction ◽  
Alkaline Phosphatase Isoenzymes ◽  
Agarose Electrophoresis ◽  
Steep Decline ◽  
Almost All

Abstract We separated isoenzymes of alkaline phosphatase (ALP; EC 3.1.3.1) in 1383 sera of normal individuals (ages 4-65 years) by agarose electrophoresis with the Isopal system (Analis). As expected, the predominant isoenzyme in children was of bone origin, and almost all (99%) of the children had low activities of a second bone fraction, "bone variant" ALP. The "bone variant" disappeared after age 17 in girls and after age 20 in boys. The highest (median) bone ALP activity was reached at age 9 to 10 in girls and at age 13 to 14 in boys, followed by a gradual decline in girls and a steep decline in boys. During adulthood, activity of the bone fraction was constant and no significant differences were observed between sexes, neither for bone nor for liver ALP activity. The latter remained almost unchanged throughout life. We observed no high-Mr ALP activity in children, whereas sera from 60% of the adults contained low activities of high-Mr ALP. Intestinal ALP (soluble form) and "intestinal variant" ALP (hydrophobic form) were frequently present, in 21% and 37% of all samples, respectively. No significant differences were observed between age groups and sexes for the intestinal isoenzymes.

Time-dependent changes in bone, placental, intestinal, and hepatic alkaline phosphatase activities in serum during human pregnancy.

1987 ◽  
Vol 33 (10) ◽  
pp. 1801-1806 ◽  
Author(s):  
G J Valenzuela ◽  
L A Munson ◽  
N M Tarbaux ◽  
J R Farley
Keyword(s):  
Alkaline Phosphatase ◽  
Pregnant Women ◽  
Bone Alkaline Phosphatase ◽  
Premature Labor ◽  
Premature Rupture Of Membranes ◽  
Premature Rupture ◽  
Serum Samples ◽  
Lactating Women ◽  
Alp Activity ◽  
Duration Of Pregnancy

Abstract To measure changes in bone alkaline phosphatase (EC 3.1.3.1) activity in serum as a function of duration of pregnancy, we adapted our existing alkaline phosphatase (ALP) isoenzyme assay (which has been used to measure bone, hepatic, and intestinal ALP activities in serum, in the absence of placental ALP) to allow quantification of individual ALP isoenzyme activities in the presence of placental ALP. The resulting CV for repeat measurements of bone ALP activity in artificial isoenzyme mixtures ranged from 23% for samples in which the bone isoenzyme represented 7% of total ALP activity to 11% for samples in which bone ALP accounted for 48% of total ALP activity. Values for repeat determinations of bone ALP activity in human serum samples (i.e., including samples obtained from pregnant women and from nonpregnant controls) varied by an average of 18%. We find, in initial applications of this method, that (a) the amount of bone ALP activity in serum is increased during pregnancy (P less than .001), and remains increased at six weeks postpartum, in non-lactating women (P less than .001), and (b) bone ALP activity at term was not significantly different in pregnant women with pre-eclampsia, diabetes, premature rupture of membranes, or premature labor, compared with normal pregnancies at term. Our data support the hypothesis that maternal bone formation may be increased during pregnancy.

Multicenter evaluation of Iso-ALP test kit for measurement of bone alkaline phosphatase activity in serum and plasma

1993 ◽  
Vol 39 (4) ◽  
pp. 648-652 ◽  
Author(s):  
S B Rosalki ◽  
A Y Foo ◽  
A Burlina ◽  
W Prellwitz ◽  
P Stieber ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Bone Alkaline Phosphatase ◽  
Alp Activity ◽  
Test Kit ◽  
Reference Limits ◽  
Bone Fraction ◽  
Adults And Children ◽  
Wheat Germ Lectin

Abstract A test kit (Iso-ALP, Boehringer Mannheim) for measuring human bone alkaline phosphatase activity in serum or plasma was evaluated in five laboratories in three countries. The assay is based on the principle described by Rosalki and Foo (Clin Chem 1984;30:1182-6) and uses wheat germ lectin to precipitate bone alkaline phosphatase. Residual ALP in the supernate in comparison with total ALP is used to quantify the bone fraction. The imprecision of residual ALP measurement was low (median between-run CV 4.9%) and comparable with that of total ALP. Linearity of precipitation was demonstrable up to a bone ALP activity (diethanolamine buffer 37 degrees C) of 2000 U/L, though a matrix effect was observed for dilutions of high-activity sera in saline or bovine serum albumin. For assaying patients' samples, different batches of lectin demonstrated excellent comparability. Taking electrophoresis as a basis for standardization, we determined that the lectin precipitated approximately 90% of bone ALP, but < 5% of nonbone ALP. From this we derived serum/plasma upper reference limits for bone ALP activity in adults and children.

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