scholarly journals Clinical Significance of Immunoassays for Type-5 Tartrate-resistant Acid Phosphatase

1999 ◽  
Vol 45 (12) ◽  
pp. 2150-2157 ◽  
Author(s):  
Yuri R Nakasato ◽  
Anthony J Janckila ◽  
Jussi M Halleen ◽  
H Kalervo Vaananen ◽  
Stephanie P Walton ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Rheumatic Disease ◽  
Healthy Subjects ◽  
Specific Activity ◽  
Biological Significance ◽  
Tartrate Resistant Acid Phosphatase ◽  
Endstage Renal Disease ◽  
Trap Activity ◽  
Specificity And Sensitivity ◽  
The Mean

Abstract Background: Tartrate-resistant acid phosphatase (TRAP; EC 3.1.3.2) is a product of osteoclasts and a biochemical marker of bone resorption rate. However, erythrocytes and platelets contribute to total TRAP activity in serum, reducing the specificity of direct biochemical assays in serum. Osteoclast TRAP is also known as type-5 TRAP and is antigenically unique. Immunoassays are sought to improve the specificity and sensitivity of TRAP as a bone marker. Methods: We developed two colorimetric microplate assays for type-5 TRAP: an enzyme capture immunoassay to measure antibody-bound enzymatic activity, and a two-site immunoassay to measure bound enzyme protein. Both use the same monoclonal antibody (14G6) to capture type-5 TRAP, which permits determination of specific activity of serum TRAP in health and disease. Results: Both TRAP assays were linear from one-tenth to fivefold the mean value in 18 healthy subjects. In these subjects, the mean (SD) TRAP activity was 3.2 (0.54) U/L for the enzyme capture assay and 37 (13) μg/L for the two-site assay. Mean TRAP activity was not significantly increased in 64 patients with endstage renal disease requiring hemodialysis (HD) or 99 unselected patients with rheumatic diseases. By contrast, TRAP protein was increased in both the HD and rheumatic disease groups. The specific activity of TRAP in the 17 of 64 HD sera that had increased TRAP activity (0.088 U/μg) was similar to that in healthy subjects (0.091 U/μg). By contrast, the specific activity of TRAP in the 31 of 99 rheumatic sera with increased TRAP protein (0.035 U/μg) was significantly decreased. Conclusions: Wide sample distributions for TRAP activity in HD patients and TRAP protein in rheumatic disease patients suggest the presence of subpopulations of HD patients with increased TRAP activity and of rheumatic patients with increased TRAP protein. Each assay for TRAP activity and protein may have its own biological significance and clinical applications in specific groups of patients.

scholarly journals Tartrate-Resistant Acid Phosphatase 5b is a Useful Serum Marker for Diagnosis and Recurrence Detection of Giant Cell Tumor of Bone

2012 ◽  
Vol 6 (1) ◽  
pp. 392-399 ◽  
Author(s):  
Tetsuya Shinozaki ◽  
Kenichi Saito ◽  
Tsutomu Kobayashi ◽  
Takashi Yanagawa ◽  
Kenji Takagishi
Keyword(s):  
Acid Phosphatase ◽  
Local Recurrence ◽  
Mean Value ◽  
Cell Tumor ◽  
Control Group ◽  
Primary Group ◽  
Tartrate Resistant Acid Phosphatase ◽  
Tracp 5B ◽  
The Mean ◽  
Recurrence Group

Serum tartrate-resistant acid phosphatase (TRACP) 5b was investigated for use as a marker for diagnosis of giant cell tumor (GCT) of bone and for detection of its recurrence.Four patients with GCT of bone who were initially referred to our hospital were classified as a primary group. Three patients who had local recurrence following curettage were classified as a local recurrence group. Five with no recurrence were classified as a no-recurrence group. Eighteen patients with primary and metastatic malignant bone tumors were also enrolled in the study as a control group. Serum TRACP 5b was measured before the biopsy in all patients and was measured periodically after the operation in patients with GCT of bone. Studentt-tests were used for statistical analyses.TRACP 5b was greater than 1500 Um/dL in all primary group patients. Mean TRACP 5b values decreased gradually with post-operative time, showing lower values until local recurrence. The mean value of TRACP 5b of the local recurrence group (753 ± 68.7 mU/dL) was significantly higher than that of the no-recurrence group (340.6 ± 78.3 mU/dL). The mean value of TRACP 5b of the control group (466.9 ± 130.3 mU/dL) was much lower than that of the primary group and markedly lower than that of the local recurrence group. However, no significant difference was found between the no-recurrence group and the control group.Serum TRACP 5b is a useful and convenient marker for diagnosing GCT of bone and for predicting its recurrence.

scholarly journals Development of a Kinetic Assay for Band 5b Tartrate-resistant Acid Phosphatase Activity in Serum

2000 ◽  
Vol 46 (4) ◽  
pp. 469-473 ◽  
Author(s):  
Mamoru Nakanishi ◽  
Kousei Yoh ◽  
Toshihide Miura ◽  
Tatsuya Ohasi ◽  
Shiba Kumar Rai ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Kinetic Method ◽  
Assay Method ◽  
Tartrate Resistant Acid Phosphatase ◽  
Specific Method ◽  
Serum Samples ◽  
Kinetic Assay ◽  
Specific Measurement ◽  
The Mean ◽  
Inhibited Enzyme

Abstract Background: Band 5 tartrate-resistant acid phosphatase (TrACP; EC 3.1.3.2) consists of two isoenzymes, bands 5a and 5b, of which band 5b TrACP is considered to be derived from bone. However, no kinetic method for the specific measurement of band 5b TrACP in serum is available. Our aim was to develop a kinetic assay method for the specific measurement of band 5b TrACP in serum. Methods: Band 5b TrACP was measured kinetically in serum as tartrate-resistant fluoride-sensitive heparin-resistant ACP with 2,6-dichloro-4-acetylphenyl phosphate as substrate at pH 6.6. Results: Heparin inhibited band 5a TrACP but had no effect on band 5b TrACP in serum or in bone extract. The presence of EDTA or ascorbic acid had no effect, but dithiothreitol inhibited enzyme activity. The within-run (n = 20) and between-run (n = 20) CVs of band 5b TrACP activity were 3.3–5.8% and 5.0–7.3%, respectively. The mean ± SD values of band 5b TrACP activity in males (n = 25) and females (n = 57) 20–29 years of age by this method were 8.0 ± 2.2 U/L and 6.4 ± 1.8 U/L, respectively. The band 5b TrACP value was significantly higher in females >50 years of age compared with the younger subjects (20–29 years). The highest band 5b TrACP values were among children younger than 15 years. Conclusions: This kinetic assay is a simple and specific method for the measurement of band 5b TrACP in serum samples and is useful in the evaluation of bone turnover activity.

Generation and characterization of monoclonal antibodies to human type-5 tartrate-resistant acid phosphatase: development of a specific immunoassay of the isoenzyme in serum

1995 ◽  
Vol 41 (10) ◽  
pp. 1495-1499 ◽  
Author(s):  
P Chamberlain ◽  
J Compston ◽  
T M Cox ◽  
A R Hayman ◽  
R C Imrie ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Acid Phosphatase ◽  
Premenopausal Women ◽  
Tartrate Resistant Acid Phosphatase ◽  
Time Resolved ◽  
Specific Serum ◽  
Serum Trap ◽  
Human Osteoclast ◽  
The Mean

Abstract We have characterized four monoclonal antibodies (mAbs) to the purple ("tartrate-resistant," band 5) acid phosphatase of the human osteoclast (TRAP) and used these to develop a specific serum immunoassay. All four mAbs are of high affinity (Ka = 1-5 x 10(8) L/mol) with a very fast Kassoc (0.2-2.0 x 10(5) L mol-1 s-1) and a moderate Kdissoc (1-3 x 10(-3) s). Two of the mAbs were selected to develop a time-resolved fluorescence immunoassay to measure serum concentrations of TRAP. The mean serum immunoreactive TRAP in a group of healthy premenopausal women and men was 3.7 +/- 1.8 micrograms/L (mean +/- SD) and 3.5 +/- 1.6 micrograms/L, respectively. Significantly higher concentrations of TRAP were found in postmenopausal women (6.3 +/- 2.3 micrograms/L) and in eight patients with Gaucher disease (19.3 +/- 4.7 micrograms/L). Further studies are required to investigate the value of serum TRAP as a marker of bone resorption.

scholarly journals Purification and characterization of a tartrate-resistant acid phosphatase from human osteoclastomas

1989 ◽  
Vol 261 (2) ◽  
pp. 601-609 ◽  
Author(s):  
A R Hayman ◽  
M J Warburton ◽  
J A S Pringle ◽  
B Coles ◽  
T J Chambers
Keyword(s):  
Acid Phosphatase ◽  
Gel Electrophoresis ◽  
Biochemical Characterization ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Tartrate Resistant Acid Phosphatase ◽  
Bovine Bone ◽  
Ph Optimum ◽  
Original Tumour

Tartrate-resistant acid phosphatase is one of the major enzymes produced and secreted by osteoclasts. To obtain sufficient enzyme for biochemical characterization, we have purified this enzyme from human osteoclastomas by sequential chromatography on SP-Sephadex, CM-Sephadex, hydroxylapatite, Sephadex G-150 and concanavalin A-Sepharose. The purification over the original tumour extract was about 2000-fold, with a yield of 10%. The enzyme appeared to be homogeneous when assessed by SDS/polyacrylamide-gel electrophoresis. Both gel filtration and SDS/polyacrylamide-gel electrophoresis indicated an Mr of about 30,000. The reduced and alkylated enzyme consists of two subunits with Mrs of 15,000 and 17,500. The N-terminal amino acid sequence of both subunits indicates that there is a high degree of identity between the osteoclastoma enzyme and similar enzymes purified from spleen and uterus. Using 4-methylumbelliferyl phosphate as substrate, the specific activity of the purified enzyme was 387 units.mg-1, and the Km was 284 microns. The pH optimum was 5.7. Unlike similar enzymes purified from human and bovine bone, osteoclastoma acid phosphatase is not activated by reducing agents (2-mercaptoethanol or ascorbic acid). The enzyme contains 4.8 mol of Fe2+/3+, 0.3 mol of Mn2+ and 1.7 mol of Mg2+ per mol of enzyme. Although the enzyme loses 50% of its activity in the presence of EDTA, it is not inhibited by the iron chelator 1,10-phenanthroline. However, the enzyme is activated to a small extent by Mn2+ and Mg2+. Using a variety of substrates and inhibitors, we demonstrate that there are differences between the osteoclastoma acid phosphatase and the enzyme purified from other sources.

Tartrate-resistant acid phosphatase (trap) activity as osteoclastic marker: Comparison between cytochemical assessment and serum assay

1996 ◽  
Vol 6 (S1) ◽  
pp. 168-168 ◽  
Author(s):  
P. Ballanti ◽  
S. Minisola ◽  
M. T. Pacitti ◽  
L. Scarnecchia ◽  
R. Rosso ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Tartrate Resistant Acid Phosphatase ◽  
Trap Activity ◽  
Serum Assay ◽  
Marker Comparison

scholarly journals Tartrate-resistant acid phosphatase as a biomarker of bone turnover in dog

2011 ◽  
Vol 63 (1) ◽  
pp. 40-45 ◽  
Author(s):  
C.P Sousa ◽  
F Nery ◽  
J.T Azevedo ◽  
C.A Viegas ◽  
M.E Gomes ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Individual Variation ◽  
Bone Alkaline Phosphatase ◽  
Tartrate Resistant Acid Phosphatase ◽  
Biological Variability ◽  
Trap Activity ◽  
Non Invasive ◽  
Serum Minerals ◽  
Serum Trap

Values of serum tartrate-resistant acid phosphatase ( TRAP) activity were obtained in adult dogs and its biological variability was assessed. Nine healthy skeletally mature Portuguese Podengo dogs were used for the determination of TRAP, total and bone alkaline phosphatase serum activities, and also to study their relationship with serum minerals, namely calcium (Ca), phosphorous (P), and magnesium (Mg). The serum TRAP activity was 2.19±0.56IU/mL, with intra-individual variation of 18.3% and inter-individual variation of 25.6%. Significant correlations were observed between serum TRAP activity and Ca (r=-0.3431; P<0.05), Ca and Mg (r=-0.787; P<0.01), and TRAP and Mg (r=0.397; P<0.05). The results indicate that serum TRAP activity in dog could be of great value in research and in clinical practice, providing complementary non-invasive information on bone metabolism

Tartrate-resistant Acid Phosphatase (TRAP) Activity in Serum: Potential Use in Assessing Bone Resorption in Patients with Multiple Myeloma

1990 ◽  
Vol 5 (2) ◽  
pp. 61-64 ◽  
Author(s):  
M. Monti ◽  
A. Scazzoso ◽  
G. Calzaferri ◽  
I. Santi ◽  
E. D'Aprile ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Acid Phosphatase ◽  
Bone Resorption ◽  
Bone Lesions ◽  
Tartrate Resistant Acid Phosphatase ◽  
Trap Activity ◽  
Serum Trap ◽  
The Difference ◽  
Young Subjects ◽  
Hydroxyproline Excretion

We measured serum tartrate-resistant acid phosphatase (TRAP) activity in 120 healthy subjects and 35 patients with multiple myeloma as well as urinary hydroxyproline excretion in the myeloma patients. Young subjects (0-18 years) showed higher TRAP levels (ANOVA p < 0.01) compared with the other age classes due to the more active bone remodelling processes associated with growth. Myeloma patients with bone lytic lesions (MM+) showed higher serum TRAP values than controls (p < 0.01). Hydroxyproline excretion was higher in MM + patients but the difference between patients with and without bone lesions was not statistically significant. Our data suggest that serum TRAP activity may be a suitable, simple biochemical test to assess bone turnover in patients with multiple myeloma but that its clinical usefulness as a marker of bone resorption needs further evaluation.

scholarly journals Improved method for measuring tartrate-resistant acid phosphatase activity in serum

1998 ◽  
Vol 44 (2) ◽  
pp. 221-225 ◽  
Author(s):  
Mamoru Nakanishi ◽  
Kousei Yoh ◽  
Kagehiro Uchida ◽  
Souji Maruo ◽  
Akira Matsuoka
Keyword(s):  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Elderly Subjects ◽  
Tartrate Resistant Acid Phosphatase ◽  
Kinetic Measurement ◽  
Improved Method ◽  
Hexadimethrine Bromide ◽  
The Mean

Abstract We describe an improved method for the kinetic measurement of tartrate-resistant acid phosphatase (TrACP; EC 3.1.3.2) activity in serum. Of the TrACP derived from erythrocytes, platelets, and macrophages (osteoclasts and others), that from the first two sources is also resistant to fluoride, whereas skeletal TrACP is sensitive to fluoride. Thus, osteoclast-derived TrACP can be measured specifically by exploiting its sensitivity to fluoride. We measured the activity of tartrate-resistant and fluoride-sensitive acid phosphatase (TrFsACP) by using 2,6-dichloro-4-acetylphenyl phosphate as substrate at pH 6.2. The activity of TrFsACP in serum was increased by adding hexadimethrine bromide (Polybrene) to the reaction mixture. This method was not influenced by hemolysis with hemoglobin concentrations as great as 0.9 g/L. The mean ± SD values of TrFsACP activity by this method were 20.4 ± 2.8 and 16.4 ± 2.3 U/L for young (ages 20–29 years) men (n = 34) and women (n = 50), respectively. The highest mean TrFsACP activity was found among children younger than 15 years, followed by that in elderly subjects (older than 60).

scholarly journals Tartrate-resistant Acid Phosphatase Isoform 5b as Serum Marker for Osteoclastic Activity

2001 ◽  
Vol 47 (1) ◽  
pp. 74-80 ◽  
Author(s):  
Anthony J Janckila ◽  
Karen Takahashi ◽  
Susan Z Sun ◽  
Lung T Yam
Keyword(s):  
Acid Phosphatase ◽  
Type I Collagen ◽  
Bone Alkaline Phosphatase ◽  
Serum Marker ◽  
Tartrate Resistant Acid Phosphatase ◽  
Type I ◽  
Enzyme Protein ◽  
Endstage Renal Disease ◽  
Osteoclastic Activity ◽  
Tracp 5B

Abstract Background: Tartrate-resistant acid phosphatase (AcP) 5b is a marker of osteoclastic activity and bone resorption. Immunoassays for serum TRAcP may lack sensitivity and specificity because of the presence of non-bone isoform 5a. The purpose of this study was to isolate the serum isoforms, quantify their disease-related expressions, and test an improved immunoassay for TRAcP 5b. Methods: We separated TRAcP isoforms chromatographically from pooled sera of healthy, rheumatoid arthritis (RA) and endstage renal disease (ESRD) subjects. TRAcP isoforms were identified by electrophoresis and quantified by biochemical and immunochemical assays. Serum TRAcP activity in healthy, RA, and ESRD cohorts was assessed at pH 5.5 and 6.1, and compared with bone alkaline phosphatase (BAP) and N-telopeptides of type I collagen (NTx). Results: TRAcP isoforms 5a and 5b were present in all sera; 5b was identical to osteoclastic TRAcP. In serum from healthy subjects, 5a accounted for 87% of the enzyme protein but only 55% of the activity. In RA, both isoforms were increased two- to threefold in protein, but their specific activities were subnormal. In ESRD, only 5b was abnormal, being increased fivefold in protein and threefold in activity. In RA sera, TRAcP activity did not correlate with either BAP or NTx. In ESRD sera, TRAcP activity correlated with BAP and NTx only when measured at pH 6.1. Conclusions: All sera contained both TRAcP isoforms 5a and 5b, but only 5b was present in bone. TRAcP isoform expression was variable in different diseases. Measurement of TRAcP activity at pH 6.1 improves the specificity of immunoassay for isoform 5b.

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