Polydopamine-mediated immobilization of phenylboronic acid on magnetic microspheres for selective enrichment of glycoproteins and glycopeptides
作者:Jiangnan Zheng、Zian Lin、Lan Zhang、Huanghao Yang
DOI:10.1007/s11426-014-5286-5
日期:2015.6
In this work, we demonstrate for the first time, a method to synthesize phenylboronic acid-Fe3O4@polydopamine (Fe3O4@PDA-PBA) magnetic microspheres via the combination of mussel-inspired polydopamine coating and click chemistry. Uniform-size and core-shell structured Fe3O4@PDA-PBA magnetic microspheres with a core diameter of ∼240 nm and a shell thickness of ∼13 nm were obtained as identified by the characterization of the morphology, structure and composition of the synthesized microspheres. We evaluated the selectivity and binding capacity of the Fe3O4@PDA-PBA magnetic microspheres by using standard glycoproteins (ovalbumin, immunoglobulin G and catalase) and nonglycoproteins (human serum albumin, bovine hemoglobin, myoglobin, lysozyme, and ribonuclease A) as model proteins. Adsorption experiments, SDS-PAGE and mass spectrometry analysis demonstrated that the Fe3O4@PDA-PBA magnetic microspheres had much high binding capacity and selectivity for glycoproteins/glycopeptides compared to nonglycoproteins/nonglycopeptides. In addition, the practicability of the Fe3O4@PDA-PBA magnetic microspheres was further assessed by selective capture of glycoproteins from healthy human serum. The good results demonstrated its potential in glycoproteome analysis.
在这项工作中,我们首次展示了一种结合贻贝启发的聚多巴胺涂层和点击化学合成苯硼酸-Fe3O4@聚多巴胺(Fe3O4@PDA-PBA)磁性微球的方法。通过形态、结构和组成的表征,我们得到了尺寸均匀、核壳结构的Fe3O4@PDA-PBA磁性微球,其核直径约为240 nm,壳厚度约为13 nm。我们评估了Fe3O4@PDA-PBA磁性微球的特异性和结合能力,使用标准糖蛋白(卵清蛋白、免疫球蛋白G和过氧化氢酶)和非糖蛋白(人血清白蛋白、牛血红蛋白、肌红蛋白、溶菌酶和核糖核酸酶A)作为模型蛋白。吸附实验、SDS-PAGE和质谱分析表明,与非糖蛋白/非糖肽相比,Fe3O4@PDA-PBA磁性微球对糖蛋白/糖肽具有更高的结合能力和特异性。此外,我们还通过从健康人血清中选择性捕获糖蛋白,进一步评估了Fe3O4@PDA-PBA磁性微球的实用性。得到的结果表明,其在糖蛋白组分析中具有潜在的应用价值。