Liu Yujie, Fu Dongmei, Yu Long, Xiao Yuansheng, Peng Xiaojun, Liang Xinmiao
State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116023, China; Key Lab of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
Key Lab of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
J Chromatogr A. 2016 Jul 15;1455:147-155. doi: 10.1016/j.chroma.2016.05.093. Epub 2016 May 27.
Lectin affinity chromatography (LAC) is an important enrichment technique in glycoproteomics analysis. In order to improve the effectiveness of enrichment, it is necessary to develop LAC materials with high specificity and efficiency. Herein, using oxidized dextran as the spacer, a silica-based concanavalin A material (SiO2-ODex Con A) was synthesized to enrich glycoproteins/glycopeptides. For comparison, the SiO2-Ald Con A material was synthesized using conventional (3-glycidoxypropyl) triethoxysilane (GPMS) as the initial spacer arm. The analytical merits of both Con A materials, such as non-specific adsorption, binding capacity and trapping efficiency, have been evaluated using ovalbumin. Under high performance liquid affinity chromatography (HPLAC) mode, the SiO2-ODex Con A material was highly effective in the enrichment of glycoproteins/glycopeptides attached to high-mannose-type and bi-antennary complex-type glycans. The promising potential of the SiO2-ODex Con A material was demonstrated by selective fractionation of glycoproteins from complex biological samples for glycosylation analysis.
凝集素亲和色谱法(LAC)是糖蛋白质组学分析中的一种重要富集技术。为了提高富集效果,有必要开发具有高特异性和高效率的LAC材料。在此,以氧化葡聚糖为间隔臂,合成了一种基于二氧化硅的伴刀豆球蛋白A材料(SiO2-ODex Con A)用于富集糖蛋白/糖肽。为作比较,使用传统的(3-缩水甘油氧基丙基)三乙氧基硅烷(GPMS)作为初始间隔臂合成了SiO2-Ald Con A材料。使用卵清蛋白评估了两种伴刀豆球蛋白A材料的分析优点,如非特异性吸附、结合能力和捕获效率。在高效液相亲和色谱(HPLAC)模式下,SiO2-ODex Con A材料在富集连接有高甘露糖型和双天线复合型聚糖的糖蛋白/糖肽方面非常有效。通过从复杂生物样品中选择性分级分离糖蛋白用于糖基化分析,证明了SiO2-ODex Con A材料具有广阔的应用前景。