DETECTION AND MOLECULAR-WEIGHT DETERMINATION OF POLYETHYLENE GLYCOL-MODIFIED HIRUDIN BY STAINING AFTER SODIUM DODECYL-SULFATE POLYACRYLAMIDE-GEL ELECTROPHORESIS
DETECTION AND MOLECULAR-WEIGHT DETERMINATION OF POLYETHYLENE GLYCOL-MODIFIED HIRUDIN BY STAINING AFTER SODIUM DODECYL-SULFATE POLYACRYLAMIDE-GEL ELECTROPHORESIS
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DOI：
10.1016/0003-2697(92)90460-o
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发表时间：
1992-02-01
影响因子：
2.9
通讯作者：
KURFURST, MM
中科院分区：
文献类型：
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作者：
KURFURST, MM
本文描述了一种在SDS-PAGE后直接检测聚乙二醇化蛋白的通用方法。用碘化钡溶液对聚乙二醇（PEG）分子附着的蛋白质进行染色。染色是基于碘化钡与PEG复合物的形成。所述方法将PEG分子的特异性染色与SDS-PAGE方法的高分辨率相结合。结果表明，在SDS-PAGE和IEF上可检测到聚乙二醇化蛋白，其浓度为考马斯蛋白染色所检测不到的浓度。本文还介绍了用不同分子量的聚乙二醇校准SDS-PAGE测定聚乙二醇化水蛭素的分子量。在所用条件下，PEG在电泳过程中显示线性迁移率。然而，使用非聚乙二醇化蛋白质作为标准品导致不正确的分子量值，因为聚乙二醇化蛋白质在电泳过程中的迁移率较低。所描述的方法可能反映了用于测定聚乙二醇化蛋白质的分子量的一般方法。
This article describes a general method for detecting pegylated proteins directly after SDS-PAGE. The proteins to which polyethylene glycol (PEG) molecules are attached are stained with a barium iodide solution. The staining is based on the formation of a barium iodide complex with PEG. The described method combines a specific staining of PEG molecules with the high resolution of the SDS-PAGE method. It is shown that pegylated protein is detectable on SDS-PAGE as well as on IEF at concentrations that are not detectable by Coomassie protein staining. This paper also describes the determination of the molecular weight of pegylated hirudin by calibrating SDS-PAGE with polyethylene glycol of different molecular weight. Under the conditions used, PEG showed linear mobility during electrophoresis. However, the use of nonpegylated proteins as standards resulted in incorrect molecular weight values due to the lower mobility of the pegylated protein during electrophoresis. The method described might reflect a general method for determining molecular weight of pegylated proteins.