College of Pharmacy, State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin 300071, People's Republic of China.
Biomacromolecules. 2013 Jan 14;14(1):124-31. doi: 10.1021/bm301477z. Epub 2012 Dec 18.
Functionalized alkynyl polyvinyl alcohol magnetic microspheres (PVA MMs) were developed for the specific enrichment of sialic acid-rich glycoproteins by click chemistry. The capture capability for proteins was evaluated through a novel dual-labeled bovine serum albumin (BSA) that utilizes fluorescence resonance energy transfer (FRET). The PVA MM parameters, including the size and coverage of functionalized groups, were optimized by response surface methodology. The optimal parameters obtained were 1.25-6.31 μm in size and 48.53-73.05% in coverage. Then, the optimal PVA MMs were synthesized, and the morphology and surface chemical properties were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FT-IR). To capture glycoproteins from the cell surface, a bioorthogonal chemical method was applied to metabolically label them with an azide group. The functionalized alkynyl PVA MMs showed a high specificity and strong binding capability for glycoproteins through a [3 + 2] cycloaddition reaction. The results indicated that the functionalized alkynyl PVA MMs could be applied to the enrichment of cell glycoproteins, and the merits of the MMs suggested an attractive and potential way to facilitate glycoprotein research.
功能化炔基聚乙烯醇磁性微球（PVA MM）通过点击化学被开发用于特异性富集富含唾液酸的糖蛋白。通过利用荧光共振能量转移（FRET）的新型双标记牛血清白蛋白（BSA）评估了蛋白质的捕获能力。通过响应面法优化了 PVA MM 的参数，包括功能化基团的大小和覆盖率。获得的最佳参数为 1.25-6.31μm 的大小和 48.53-73.05%的覆盖率。然后，合成了最佳的 PVA MM，并通过透射电子显微镜（TEM）、扫描电子显微镜（SEM）和傅里叶变换红外光谱（FT-IR）对其形态和表面化学性质进行了表征。为了从细胞表面捕获糖蛋白，应用生物正交化学方法将其用叠氮基团进行代谢标记。通过[3+2]环加成反应，功能化炔基 PVA MM 对糖蛋白表现出高特异性和强结合能力。结果表明，功能化炔基 PVA MM 可用于细胞糖蛋白的富集，MM 的优点为糖蛋白研究提供了一种有吸引力和有潜力的方法。
