Proteomics Laboratory, Medical and Healthy Analytical Center, Peking University, Beijing 100191, China.
Carbohydr Polym. 2012 Oct 1;90(2):799-804. doi: 10.1016/j.carbpol.2012.05.090. Epub 2012 Jun 16.
In this study we describe a method for highly specific enrichment of glycopeptides with boronic acid-functionalized chitosan polymeric nanospheres and matrix assisted laser desorption-ionization mass spectrometry (MALDI-MS). This is the first time chitosan has been used to create nanosphere support material for selective enrichment of glycopeptides by modification with glycidyl methacrylate (GMA) and derivatization with 3-aminophenylboronic acid (APB). Due to their multifunctional chemical moieties, these 20-100 nm chitosan-GMA-APB nanospheres have unique properties, such as good dispersibility, good biocompatibility and chemical stability, as well as augmented specificity with glycopeptides. Enrichment conditions were optimized by using trypsin digested glycoprotein horseradish peroxidase. The high specificity of chitosan-GMA-APB nanospheres was demonstrated by effectively enriching glycopeptides from a digest mixture of horseradish peroxidase and nonglycoproteins (bovine serum albumin (BSA)).
在这项研究中，我们描述了一种使用硼酸功能化壳聚糖聚合纳米球和基质辅助激光解吸电离质谱（MALDI-MS）高度特异性富集糖肽的方法。这是壳聚糖首次被用于通过与甲基丙烯酸缩水甘油酯（GMA）进行修饰以及与 3-氨苯基硼酸（APB）进行衍生化来创建用于选择性富集糖肽的纳米球支撑材料。由于其多功能化学部分，这些 20-100nm 的壳聚糖-GMA-APB 纳米球具有独特的性质，例如良好的分散性、良好的生物相容性和化学稳定性，以及与糖肽增强的特异性。通过使用胰蛋白酶消化糖蛋白辣根过氧化物酶优化了富集条件。壳聚糖-GMA-APB 纳米球的高特异性通过有效地从辣根过氧化物酶和非糖蛋白（牛血清白蛋白（BSA））的消化混合物中富集糖肽得到了证明。
