用SDS-填充胶毛细管电泳法测定蛋白质分子量

Determination of Molecular Weight of Proteins by SDS Gel-filled Capillary Electrophoresis

目的： 建立蛋白质分子量测定的毛细管电泳方法。方法： 以葡聚糖为分离介质， 线性聚丙烯酰胺涂渍毛细管柱为支持介质， 柱上２１４ｎｍ 检测， 进行蛋白质分离。结果： 被分离的蛋白质分子量的对数与其相对迁移率之间具有良好的线性关系（ｒ ＝ ０－９９７） ； 涂渍毛细管柱可以使用多达１００次而未出现分离效率的降低； 同一毛细管柱和不同毛细管柱迁移时间的 ＲＳＤ 分别小于１－０ ％ 和１－３ ％ ； 用十二烷基硫酸钠－ 毛细管胶电泳（ＳＤＳ－ ＣＧＥ） 方法测定的１５ 种蛋白质的分子量与用平板ＳＤＳ－ 聚丙烯酰胺胶电泳（ＳＤＳ－ ＰＡＧＥ） 测定的分子量具有良好的一致性。结论： 毛细管电泳法可用于蛋白质分子的分子量测定， 它具有准确、快速等优点。

Objective:To establish a capillary electrophoresis method for the determination of molecular weight of proteins.Methods:On linear polyacrylamide coated capillary column,using dextran as sieving medium,proteins were separated with on-column UV monitoring at 214 nm.Results:There is a good linear relationship between the logarithm of molecular weight and rate of relative migration of proteins( r =0 997).Coated capillary column can be used for more than 100 sample injections without loss in performance efficiency. Relative standard deviation of migration time in one capillary and different capillaries are less than 1.0% and 1.3%,respectively.The molecular weights of fifteen proteins determined by SDS capillary gel electrophoresis (SDS-CGE) are in good agreement with those obtained from the slab SDS polyacrylamide gel electrophoresis (SDS-PAGE). Conclusion: CE would be an accurate and rapid method for the determination of molecular weight of proteins.