Rosalki S B, Foo A Y
Clin Chem. 1984 Jul;30(7):1182-6.
We describe two new methods for the separation and quantification of the bone and liver isoenzymes of alkaline phosphatase (EC 3.1.3.1) in plasma. In the first, we use wheat-germ lectin to precipitate the bone isoenzyme. About 80% of this, but minimal liver isoenzyme, is precipitated. The activity of the bone isoenzyme is calculated from measuring the alkaline phosphatase activity in the precipitate, that of liver alkaline phosphatase by subtracting the activity of the bone isoenzyme from total alkaline phosphatase activity. The liver fraction will also contain biliary, intestinal, and placental alkaline phosphatase if these are present in the original plasma, but correction for such activity is readily made. In the second method, samples are separated on cellulose acetate membranes that, before electrophoresis, have been soaked in buffer containing wheat-germ lectin. The bone isoenzyme is retarded and clearly separated from the liver fraction, allowing these isoenzymes to be quantified by densitometry. Both methods are rapid, reproducible, and suitable for use in the diagnostic laboratory.
我们描述了两种用于分离和定量血浆中碱性磷酸酶（EC 3.1.3.1）骨同工酶和肝同工酶的新方法。第一种方法，我们使用麦胚凝集素沉淀骨同工酶。约80%的骨同工酶会沉淀下来，而肝同工酶沉淀极少。骨同工酶的活性通过测量沉淀物中的碱性磷酸酶活性来计算，肝碱性磷酸酶的活性则通过从总碱性磷酸酶活性中减去骨同工酶的活性来计算。如果原始血浆中存在胆汁、肠道和胎盘碱性磷酸酶，肝部分也会包含这些，但对此类活性的校正很容易进行。在第二种方法中，样品在醋酸纤维素膜上进行分离，在电泳前，醋酸纤维素膜已浸泡在含有麦胚凝集素的缓冲液中。骨同工酶迁移受阻，并与肝部分清晰分离，从而可通过光密度测定法定量这些同工酶。这两种方法都快速、可重复，适用于诊断实验室。
