玻璃微流控芯片十二烷基硫酸钠无胶筛分电泳测定免疫球蛋白片段分子量

通过实验优化了葡聚糖筛分介质和运行缓冲溶液的浓度,采用十二烷基硫酸钠(SDS)无胶筛分电泳分离体系(10%(w/v)葡聚糖,0.1%SDS,10%甘油,0.2mol/LTris-硼砂,pH8.3的缓冲液)在自制的玻璃微流控芯片上高效分离了BODIPY衍生的蛋白质分子量标准样品,连续6次电泳所得迁移时间的相对标准偏差均小于0.50%。以6种蛋白质分子量的对数对其迁移时间作图,线性回归良好(r=0.994)。采用该芯片电泳分析体系对免疫球蛋白G不同片段的分子量进行了测定,所得结果与实际基本相符。

Glass Microfluidic Chip-based Sodium Dodecyl Sulfate-Nongel Sieving Electrophoresis for Determining Molecular Weights of Immunoglobulin Fragments

The concentrations of dextran sieving matrix and running buffer were optimized.Under the optimized conditions(10%(w/v)dextran,0.1%SDS,10%glycerol,0.2 mol/L Tris-boric acid,pH 8.3),six standard proteins could be efficiently separated using sodium dodecyl sulfate(SDS)-nongel sieving electrophoresis on home-made glass microfluidic chip.The relative standard deviations(RSDs)for migration time were lower than 0.50%.A good linear relationship(r=0.994)was built between the migration time of each protein and the logarithm of its molecular weight(logMr).Using the chip-based electrophoretic separation system,the molecular weights of different fragments of immunoglobulin G after different treatments were measured,and the results matched the facts.