Benzoboroxole-functionalized magnetic core/shell microspheres for highly specific enrichment of glycoproteins under physiological conditions.
Benzoboroxole-functionalized magnetic core/shell microspheres for highly specific enrichment of glycoproteins under physiological conditions.
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DOI：
10.1002/smll.201302841
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发表时间：
2014-04
期刊：
影响因子：
13.3
通讯作者：
Y. Zhang;Wanfu Ma;D. Li;Meng Yu;Jia Guo;Changchun Wang
中科院分区：
文献类型：
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作者：
Y. Zhang;Wanfu Ma;D. Li;Meng Yu;Jia Guo;Changchun Wang
从复杂的生物样品中高效富集特异性糖蛋白对于发现生物系统中的疾病生物标志物具有重要意义。近年来，基于苯基硼酸的功能材料已被广泛用于糖蛋白的富集。然而，这种富集主要在碱性条件下进行，这与糖蛋白在中性生理条件下的状态不同，并且可能导致一些不可预测的降解。在这项研究中，按需中性富集的糖蛋白从粗生物样品中完成利用之间的可逆相互作用的顺式二醇的糖蛋白和苯并硼氧杂环戊二烯功能化的磁性复合微球（Fe 3 O 4/PAA-AOPB）。设计并构建了以高磁响应磁性超微粒（MSP）为核、表面结合大量苯并硼杂环戊二烯官能团的交联聚丙烯酸（PAA）为壳的Fe 3 O 4/PAA-AOPB复合微球。这些纳米复合材料具有富集容量大（93.9mg/g，蛋白/珠）、非特异性吸附低、富集过程快（10 min）、磁分离速度快（20 s）、回收率高等优点。此外，所制备的Fe 3 O 4/PAA-AOPB微球在大肠杆菌中对糖蛋白也表现出较高的选择性。大肠杆菌裂解物或胎牛血清中的糖蛋白，显示了巨大的潜力，鉴定低丰度的糖蛋白作为生物标志物在真实的复杂的生物系统的临床诊断。
Efficient enrichment of specific glycoproteins from complex biological samples is of great importance towards the discovery of disease biomarkers in biological systems. Recently, phenylboronic acid-based functional materials have been widely used for enrichment of glycoproteins. However, such enrichment was mainly carried out under alkaline conditions, which is different to the status of glycoproteins in neutral physiological conditions and may cause some unpredictable degradation. In this study, on-demand neutral enrichment of glycoproteins from crude biological samples is accomplished by utilizing the reversible interaction between the cis-diols of glycoproteins and benzoboroxole-functionalized magnetic composite microspheres (Fe3O4/PAA-AOPB). The Fe3O4/PAA-AOPB composite microspheres are deliberately designed and constructed with a high-magnetic-response magnetic supraparticle (MSP) core and a crosslinked poly(acrylic acid) (PAA) shell anchoring abundant benzoboroxole functional groups on the surface. These nanocomposites possessed many merits, such as large enrichment capacity (93.9 mg/g, protein/beads), low non-specific adsorption, quick enrichment process (10 min) and magnetic separation speed (20 s), and high recovery efficiency. Furthermore, the as-prepared Fe3O4/PAA-AOPB microspheres display high selectivity to glycoproteins even in the E. coli lysate or fetal bovine serum, showing great potential in the identify of low-abundance glycoproteins as biomarkers in real complex biological systems for clinical diagnoses.