Nakanishi M, Yoh K, Miura T, Ohasi T, Rai S K, Uchida K
FALCO Biosystems Co., 17-1 Nishiarami Tai, Kumiyama-cho, Kuze-gun, Kyoto 613-0036, Japan.
Clin Chem. 2000 Apr;46(4):469-73.
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.
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.
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.
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.
5型抗酒石酸酸性磷酸酶（TrACP；EC 3.1.3.2）由两种同工酶组成，即5a和5b条带，其中5b条带TrACP被认为来源于骨骼。然而，目前尚无特异性检测血清中5b条带TrACP的动力学方法。我们的目的是开发一种特异性检测血清中5b条带TrACP的动力学检测方法。
以2,6-二氯-4-乙酰苯基磷酸为底物，在pH 6.6条件下，对血清中的5b条带TrACP进行动力学检测，作为抗酒石酸、氟敏感、肝素抵抗的酸性磷酸酶。
肝素可抑制血清或骨提取物中的5a条带TrACP，但对5b条带TrACP无影响。乙二胺四乙酸（EDTA）或抗坏血酸的存在无影响，但二硫苏糖醇可抑制酶活性。5b条带TrACP活性的批内（n = 20）和批间（n = 20）变异系数分别为3.3%-5.8%和5.0%-7.3%。用该方法检测20-29岁男性（n = 25）和女性（n = 57）血清中5b条带TrACP活性的平均值±标准差分别为8.0±2.2 U/L和6.4±1.8 U/L。50岁以上女性的5b条带TrACP值显著高于年轻受试者（20-29岁）。5b条带TrACP值最高的是15岁以下儿童。
这种动力学检测方法是一种简单、特异的血清样本中5b条带TrACP检测方法，有助于评估骨转换活性。
