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Development of Monolithic Column Materials for the Separation and Analysis of Glycans

机译：用于分离和分析聚糖的整体柱材料的开发

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Monolithic column materials offer great advantages as chromatographic media in bioseparations and as solid-supports in biocatalysis. These single-piece porous materials have an interconnected ligament structure that limits the void volume inside the column, thus increasing the efficiency without sacrificing the permeability. The preparation of monolithic materials is easy, reproducible and has available a wide range of chemistries to utilize. Complex, heterogeneous and isobaric glycan structures require preparation methods that may include glycan release, separation and enrichment prior to a comprehensive and site-specific glycosylation analysis. Monolithic column materials aid that demand, as shown by the results reported by the research works presented in this review. These works include selective capture of glycans and glycoproteins via their interactions with lectins, boronic acids, hydrophobic, and hydrophilic/polar functional groups on monolith surfaces. It also includes immobilization of enzymes trypsin and PNGase F on monoliths to digest and deglycosylate glycoproteins and glycopeptides, respectively. The use of monolithic capillary columns for glycan separations through nano-liquid chromatography (nano-LC) and capillary electrochromatography (CEC) and coupling these columns to MS instruments to create multidimensional systems show the potential in the development of miniaturized, high-throughput and automated systems of glycan separation and analysis.
机译：整体柱材料具有巨大的优势，可作为生物分离中的色谱介质和生物催化中的固体载体。这些单件式多孔材料具有相互连接的韧带结构，可限制色谱柱内部的空隙体积，从而在不牺牲渗透性的情况下提高了效率。整体材料的制备是容易，可重现的，并具有广泛的化学用途。复杂的，异质的和同量异位的聚糖结构需要制备方法，其中可能包括在全面和位点特异性糖基化分析之前，聚糖的释放，分离和富集。整体柱材料可以满足这一需求，正如本综述中研究工作所报告的结果所表明的那样。这些工作包括通过聚糖与糖蛋白与凝集素，硼酸，疏水基团和整体表面上的亲水/极性官能团的相互作用来选择性捕获聚糖和糖蛋白。它还包括将胰蛋白酶和PNGase F固定在整料上，以分别消化和去糖基化糖蛋白和糖肽。使用整体式毛细管柱通过纳米液相色谱（nano-LC）和毛细管电色谱（CEC）进行聚糖分离，并将这些柱与MS仪器联用以创建多维系统，显示了开发小型化，高通量和自动化系统的潜力糖分离和分析系统。

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