Wolfson Centre for Applied Structural Biology, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Safra Campus, Givat Ram, Jerusalem, Israel.
Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Safra Campus, Givat Ram, Jerusalem, Israel.
Sci Rep. 2018 May 2;8(1):6907. doi: 10.1038/s41598-018-25246-6.
Multi-angle light scattering coupled with size exclusion chromatography (SEC-MALS) is a standard and common approach for characterizing protein mass, overall shape, aggregation, oligomerization, interactions and purity. The limited resolution of analytical SEC restricts in some instances the accurate analysis that can be accomplished by MALS. These include mixtures of protein populations with identical or very similar molecular masses, oligomers with poor separation and short peptides. Here we show that combining MALS with the higher resolution separation technique ion exchange (IEX-MALS) can allow precise analyses of samples that cannot be resolved by SEC-MALS. We conclude that IEX-MALS is a valuable and complementary method for protein characterization, especially for protein systems that could not be fully analyzed by SEC-MALS.
多角度光散射与尺寸排阻色谱(SEC-MALS)联用是一种用于表征蛋白质质量、整体形状、聚集、寡聚化、相互作用和纯度的标准和常用方法。分析型 SEC 的分辨率有限,在某些情况下会限制 MALS 能够完成的准确分析。这些情况包括具有相同或非常相似分子量的蛋白质群体混合物、分离效果不佳的寡聚物和短肽。在这里,我们表明将 MALS 与更高分辨率的分离技术离子交换(IEX-MALS)相结合,可以对 SEC-MALS 无法分辨的样品进行精确分析。我们得出结论,IEX-MALS 是一种有价值的、互补的蛋白质表征方法,特别是对于 SEC-MALS 无法完全分析的蛋白质体系。