National Glycoengineering Research Center, Shandong Provincial Key Laboratory of Carbohydrate Chemistry and Glycobiology, State Key Laboratory of Microbial Technology, Shandong University, Jinan, Shandong, 250100, China; Department of Chemistry and Chemical Biology, Department of Chemical and Biological Engineering, Department of Biology, Department of Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, 12180, United States.
Department of Chemistry and Chemical Biology, Department of Chemical and Biological Engineering, Department of Biology, Department of Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, 12180, United States.
Anal Chim Acta. 2017 Apr 8;961:91-99. doi: 10.1016/j.aca.2017.01.042. Epub 2017 Jan 31.
Heparin is a structurally complex, polysaccharide anticoagulant derived from livestock, primarily porcine intestinal tissues. Low molecular weight (LMW) heparins are derived through the controlled partial depolymerization of heparin. Increased manufacturing and regulatory concerns have provided the motivation for the development of more sophisticated analytical methods for determining both their structure and pedigree. A strategy, for the comprehensive comparison of parent heparins and their LMW heparin daughters, is described that relies on the analysis of monosaccharide composition, disaccharide composition, and oligosaccharide composition. Liquid chromatography-mass spectrometry is rapid, robust, and amenable to automated processing and interpretation of both top-down and bottom-up analyses. Nuclear magnetic resonance spectroscopy provides complementary top-down information on the chirality of the uronic acid residues and glucosamine substitution. Principal component analysis (PCA) was applied to the normalized abundance of oligosaccharides, calculated in the bottom-up analysis, to show parent and daughter correlation in oligosaccharide composition. Using these approaches, six pairs of parent heparins and their daughter generic enoxaparins from two different manufacturers were comprehensively analyzed. Enoxaparin is the most widely used LMW heparin and is prepared through controlled chemical β-eliminative cleavage of porcine intestinal heparin. Lovenox, the innovator version of enoxaparin marketed in the US, was analyzed as a reference for the daughter LMW heparins. The results, show similarities between LMW heparins from two different manufacturers with Lovenox, excellent lot-to-lot consistency of products from each manufacturer, and detects a correlation between each parent heparin and daughter LMW heparin.
肝素是一种结构复杂的多糖类抗凝剂，来源于牲畜，主要是猪的肠组织。低分子量肝素（LMWH）是通过肝素的受控部分解聚得到的。增加的制造和监管问题为开发更复杂的分析方法来确定其结构和谱系提供了动力。本文描述了一种用于全面比较母体肝素及其 LMW 肝素女儿的策略，该策略依赖于单糖组成、二糖组成和寡糖组成的分析。液相色谱-质谱法快速、稳健，适用于自上而下和自下而上分析的自动化处理和解释。核磁共振波谱提供了关于糖醛酸残基和葡萄糖胺取代的手性的互补自上而下信息。主成分分析（PCA）应用于自下而上分析中计算的寡糖的归一化丰度，以显示寡糖组成中母体和女儿的相关性。使用这些方法，全面分析了来自两个不同制造商的六对母体肝素及其女儿通用依诺肝素。依诺肝素是使用最广泛的 LMW 肝素，通过对猪肠肝素进行受控的化学 β 消除裂解制备。Lovenox 是依诺肝素在美国的创新版本，被分析为女儿 LMW 肝素的参考。结果表明，来自两个不同制造商的 LMW 肝素与 Lovenox 相似，每个制造商的产品批间一致性极好，并检测到每个母体肝素与其女儿 LMW 肝素之间的相关性。
