TY - JOUR
T1 - Development of a kinetic assay for band 5b tartrate-resistant acid phosphatase activity in serum
AU - Nakanishi, Mamoru
AU - Yoh, Kousei
AU - Miura, Toshihide
AU - Ohasi, Tatsuya
AU - Rai, Shiba Kumar
AU - Uchida, Kagehiro
PY - 2000
Y1 - 2000
N2 - 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. (C) 2000 American Association for Clinical Chemistry.
AB - 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. (C) 2000 American Association for Clinical Chemistry.
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U2 - 10.1093/clinchem/46.4.469
DO - 10.1093/clinchem/46.4.469
M3 - Article
C2 - 10759470
AN - SCOPUS:0034074149
VL - 46
SP - 469
EP - 473
JO - Clinical Chemistry
JF - Clinical Chemistry
SN - 0009-9147
IS - 4
ER -