Department of Biochemistry, Emory Comprehensive Glycomics Core, Emory University School of Medicine, Atlanta, GA 30322, Georgia.
Department of Biochemistry, Emory Comprehensive Glycomics Core, Emory University School of Medicine, Atlanta, GA 30322, Georgia.
Anal Biochem. 2020 Jun 15;599:113702. doi: 10.1016/j.ab.2020.113702. Epub 2020 Apr 8.
While glycoscience has become well recognized as an indispensable area in biomedical research, studies on the function of individual glycans remains a great challenge due to the lack of tools and methods. One of the greatest impediments to progress in this area is the lack of biomedically relevant complex glycans in sufficient quantity and purity for structural and functional analysis. Despite recent advances in chemoenzymatic synthesis of complex glycans, generating significant amounts of pure glycans is limited to laboratories with specialized expertise. We have previously reported the oxidative release of natural glycans (ORNG) using household bleach, which provides large quantities of biologically relevant glycans that can be a source of glycans in quantities (>mg scale) suitable for functional studies. However, the preparative scale separation of complicated glycan mixtures has not been studied due largely to the fact that gram quantities of starting glycans have not been available until now. Here we report the adoption of closed-loop, recycle HPLC to resolve closely related glycan structures, including complex glycan isomers at preparative scale (10-100 mg).
尽管糖科学已被公认为生物医学研究中不可或缺的领域，但由于缺乏工具和方法，研究单个聚糖的功能仍然是一个巨大的挑战。该领域进展的最大障碍之一是缺乏足够数量和纯度的用于结构和功能分析的具有生物相关性的复杂聚糖。尽管最近在复杂聚糖的化学酶合成方面取得了进展，但大量生成纯聚糖的能力仅限于具有专业知识的实验室。我们之前曾报道过使用家用漂白剂进行天然聚糖的氧化释放（ORNG），这提供了大量具有生物相关性的聚糖，可以作为大量（>毫克级）适合功能研究的聚糖来源。然而，由于直到现在才获得克数量的起始聚糖，因此尚未研究复杂糖混合物的制备规模分离。在这里，我们报告了闭路、循环 HPLC 的采用，以解决包括复杂糖异构体在内的密切相关的聚糖结构，在制备规模（10-100mg）下。
