Root C F, Fine J S, Clayson K J
Clin Chem. 1987 Jun;33(6):830-2.
We attempted to separate bone and liver alkaline phosphatase (EC 3.1.3.1) isoenzymes in human serum by isoelectric focusing on agarose gel. We found that in a pH 3-10 gradient the liver and bone isoenzymes focused into so many bands over a narrow pH range such that the information could not be quantified. However, when the bone isoenzyme in serum was first desialylated at 37 degrees C for a minimum of 6 h, catalyzed by neuraminidase (EC 3.2.1.18) at pH 5.8-6.0, we could detect four distinct bands with pls of 6.7, 6.8, 6.9, and 7.0. Under the same conditions, the liver isoenzyme in human serum focused into one band at pH 7.0. The multiple banding we observed for the desialylated bone isoenzyme has not been previously reported. The method is suited as a qualitative technique for detecting the bone alkaline phosphatase isoenzyme in serum.
我们尝试通过在琼脂糖凝胶上进行等电聚焦来分离人血清中的骨碱性磷酸酶和肝碱性磷酸酶（EC 3.1.3.1）同工酶。我们发现，在pH 3 - 10梯度下，肝和骨同工酶在狭窄的pH范围内聚焦成许多条带，以至于无法对这些信息进行定量分析。然而，当血清中的骨同工酶首先在37℃下由神经氨酸酶（EC 3.2.1.18）在pH 5.8 - 6.0催化脱唾液酸至少6小时后，我们能够检测到四个不同的条带，其等电点分别为6.7、6.8、6.9和7.0。在相同条件下，人血清中的肝同工酶在pH 7.0时聚焦成一条带。我们观察到的脱唾液酸骨同工酶的多条带现象此前尚未见报道。该方法适合作为检测血清中骨碱性磷酸酶同工酶的定性技术。
