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Clinical Chemistry 46: 469-473, 2000;
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(Clinical Chemistry. 2000;46:469-473.)
© 2000 American Association for Clinical Chemistry, Inc.

Articles

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

Mamoru Nakanishi1,a, Kousei Yoh2, Toshihide Miura3, Tatsuya Ohasi3, Shiba Kumar Rai4 and Kagehiro Uchida1

1 FALCO Biosystems Co., 17-1 Nishiarami Tai, Kumiyama-cho, Kuze-gun, Kyoto 613-0036, Japan.

2 Department of Orthopedic Surgery, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya, Hyogo 633-8501, Japan.

3 Nitto Boseki Co. Ltd, 1 Shiojima, Fukuhara, Koriyama, Fukusima 963-8061, Japan.

4 Department of Medical Zoology, Kobe University School of Medicine, 7-5-1 Kusuniki-cho, Chuo-ku 650-0017, Japan.
a Author for correspondence. Fax 0774-46-1101; e-mail falconpd{at}alles.or.jp

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.




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 Clin. Chem.Home page
J. M. Halleen, S. L. Alatalo, A. J. Janckila, H. W. Woitge, M. J. Seibel, and H. K. Vaananen
Serum Tartrate-resistant Acid Phosphatase 5b Is a Specific and Sensitive Marker of Bone Resorption
Clin. Chem., March 1, 2001; 47(3): 597 - 600.
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