Davey R, Bowen R, Cahill J
Biochem Int. 1983 May;6(5):643-51.
A high resolution method has been developed to separate the isoenzymes of galactosyltransferase by combining isoelectricfocusing (IEF) in 245 mm long agarose gels with a highly sensitive enzyme activity assay. The resolution and sensitivity is such that the isoenzyme pattern of 10 microliters human serum can be resolved. Using this method normal human serum was shown to contain at least 12 isoenzyme forms of galactosyltransferase, the major forms having isoelectric points of 4.33, 4.43, 4.51, 4.61, 4.74, 4.87, 4.96, 5.16 and 5.23. Part of the isoenzyme pattern complexity is due to sialylation of some isoenzymes. Alpha-lactalbumin-affinity chromatography, a method widely used in the purification of galactosyltransferase, causes a preferential purification of some of the isoenzyme forms.
已开发出一种高分辨率方法，通过将在245毫米长的琼脂糖凝胶中进行等电聚焦（IEF）与高灵敏度酶活性测定相结合，来分离半乳糖基转移酶的同工酶。其分辨率和灵敏度足以分辨10微升人血清的同工酶谱。使用该方法显示正常人血清至少含有12种半乳糖基转移酶同工酶形式，主要形式的等电点为4.33、4.43、4.51、4.61、4.74、4.87、4.96、5.16和5.23。同工酶谱部分复杂性归因于某些同工酶的唾液酸化。α-乳白蛋白亲和色谱法是广泛用于纯化半乳糖基转移酶的方法，它会优先纯化某些同工酶形式。
