Different degrees of glycosaminoglycan sulfation result in their different charge densities. The charge density differences impact their migration behavior in polyacrylamide gel electrophoresis and size exclusion chromatography, two of the most common methods for determining relative molecular masses of polysaccharides. In this study, we investigated the feasibility of using commercially available heparin oligosaccharides as calibrants for measuring the relative molecular masses of intermediates in a bioengineered heparin process that have different levels of sulfation. A size exclusion chromatography method was established that eliminates this charge density effect and allows the determination of relative molecular mass using a single calibration curve with heparin oligosaccharides calibrants. This is accomplished by overcoming the electrostatic interaction between the glycosaminoglycans and size exclusion chromatography stationary phase using high ionic strength mobile phase.

中文翻译：

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通过尺寸排阻色谱法对生物工程肝素工艺中不同硫酸化程度的糖胺聚糖进行分子质量表征

不同程度的糖胺聚糖硫酸化导致它们不同的电荷密度。电荷密度差异会影响它们在聚丙烯酰胺凝胶电泳和尺寸排阻色谱中的迁移行为，这是测定多糖相对分子质量的两种最常用方法。在这项研究中，我们研究了使用市售肝素寡糖作为校准物的可行性，用于测量具有不同硫酸化水平的生物工程肝素过程中中间体的相对分子质量。建立了一种尺寸排阻色谱方法，该方法消除了这种电荷密度效应，并允许使用带有肝素寡糖校准物的单一校准曲线来确定相对分子质量。