PAMA-Arg brush-functionalized magnetic composite nanospheres for highly effective enrichment of phosphorylated biomolecules.
PAMA-Arg brush-functionalized magnetic composite nanospheres for highly effective enrichment of phosphorylated biomolecules.
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DOI：
10.1039/c8tb00705e
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发表时间：
2018-06
期刊：
影响因子：
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通讯作者：
Bin Luo;Xiaoxi Zhou;Peipei Jiang;Qiangying Yi;Fang Lan;Yao Wu
中科院分区：
文献类型：
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作者：
Bin Luo;Xiaoxi Zhou;Peipei Jiang;Qiangying Yi;Fang Lan;Yao Wu
磷酸化生物分子的高效富集对于磷酸化蛋白质组的深入和全面分析至关重要。在亲和材料中，基于固定化金属亲和层析（IMAC）方法的材料最常用于磷酸肽富集。然而，IMAC材料中的金属阳离子损失问题仍然是一个挑战。在这项研究中，已经开发了一种新的策略，既高效富集磷酸化的生物分子和防止金属阳离子的损失，使用Fe 3 O 4/聚多巴胺/聚（2-氨基乙基甲基丙烯酸盐酸盐）/精氨酸（Fe 3 O 4/PDA/PAMA-Arg）复合纳米球。通过一种简单的方法制备的磁性复合纳米球，具有均匀的形态，高的饱和磁化强度（45 emu g-1），和快速的磁响应（在10 s内）。值得注意的是，PAMA-Arg刷上丰富的胍基和氨基基团赋予了纳米微球极高的检测灵敏度（0.2 fmol），优异的选择性（β-酪蛋白/BSA = 1：500），以及在磷酸肽分析中的高回收率（5个循环）。此外，Fe 3 O 4/PDA/PAMA-Arg纳米微球对模型蛋白混合物中的β-酪蛋白或卵清蛋白（OVA）具有特异性分离能力，对真实的生物样品（鸡蛋白色、脱脂牛奶和大鼠脑裂解液）中的磷酸化生物分子具有显著的选择性富集性能，在痕量生物检测和蛋白质组学分析中显示出巨大的潜力。
Highly effective enrichment of phosphorylated biomolecules is vital for an in depth and comprehensive phosphoproteome analysis. Among affinity materials, materials based on the immobilized metal affinity chromatography (IMAC) method are most commonly used in phosphopeptide enrichment. However, the problem of metal cation loss in IMAC materials is still a challenge. In this study, a novel strategy has been developed for both highly effective enrichment of phosphorylated biomolecules and prevention of metal cation loss using Fe3O4/polydopamine/poly(2-aminoethyl methacrylate hydrochloride)/arginine (Fe3O4/PDA/PAMA-Arg) composite nanospheres. The magnetic composite nanospheres were prepared by a facile method, possessing uniform morphology, a high saturation magnetization (45 emu g-1), and a quick magnetic response (within 10 s). Noticeably, abundant guanidyl and amino groups of the PAMA-Arg brushes endowed the nanospheres with extremely high detection sensitivity (0.2 fmol), excellent selectivity (β-casein/BSA = 1 : 500), and high recyclability (5 cycles) in phosphopeptide analysis. Furthermore, the Fe3O4/PDA/PAMA-Arg nanospheres exhibited specific separation capacity towards β-casein or ovalbumin (OVA) in the model protein mixture and remarkably selective enrichment performance for phosphorylated biomolecules in real biological samples (egg white, nonfat milk, and rat brain lysate), showing great potential for trace biological detection and proteomics analysis.