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首页> 外文期刊>RSC Advances >New cellulose-silica composite IMAC/C18 for the selective enrichment of phosphorylated molecules and the improved recovery of hydrophilic species
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New cellulose-silica composite IMAC/C18 for the selective enrichment of phosphorylated molecules and the improved recovery of hydrophilic species

机译：新型纤维素-二氧化硅复合材料IMAC / C18，可选择性富集磷酸化分子并提高亲水性物质的回收率

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Cellulose and silica are the traditional quality sorbents with a range of reported applications, particularly for hydrophilic biomolecules. With this in mind, cellulose and silica were brought into a composite form to bring their individual benefits on to a single platform. Cellulose and silica were chosen in order to obtain a completely hydrophilic composite, which was then derivatized for the selective enrichment of phosphopeptides and as a desalting material prior to mass spectrometric analysis. The composite was characterized by FT-IR, EDX and SEM. As a material for immobilized metal ion affinity chromatography (IMAC), the cellulose-silica composite enriches phosphopeptides from a complex mixture in which beta-casein is spiked in de-phosphorylated HeLa cell extract. The inherent hydrophilic nature of the composite gives a higher selectivity of up to 2000 times the complexity level and sensitivity down to 1 femtomole for phosphopeptides. As well as the immobilized metal ions on the IMAC composite, the roles of the base materials in the composite, i.e. cellulose and silica were also tested in the enrichment which helped thereof in optimizing the best sample preparation protocol. With the introduction of successive elution conditions, phospholipids and phosphopeptides were enriched and identified from egg yolk digest using a single batch extraction method. The phosphopeptides were analyzed with MALDI-MS, whereas the low molecular weight phospholipids were analysed using newly designed gold nanoparticles and carbon based materials through LDI-MS. Phosphopeptides were also enriched from complex serum digest and discussed with relevance to prostate cancer. As a reverse-phase (RP) material, it provides a combination of the hydrophilic composite and the hydrophobic C18 chain. Efficient desalting of higher salt concentrations, improved recovery of hydrophilic peptides and high sequence coverage in comparison to commercial materials makes cellulose-silica C18 a better desalting tool with in-house economical synthesis.
机译：纤维素和二氧化硅是传统的优质吸附剂，具有许多已报道的应用，特别是对于亲水性生物分子。考虑到这一点，将纤维素和二氧化硅制成复合形式，以将其各自的优势带到一个平台上。选择纤维素和二氧化硅以获得完全亲水的复合材料，然后将其衍生化以选择性富集磷酸肽，并在质谱分析之前用作脱盐材料。通过FT-IR，EDX和SEM对复合材料进行了表征。作为固定金属离子亲和色谱（IMAC）的材料，纤维素-二氧化硅复合材料从复杂混合物中富集磷酸肽，在复杂混合物中，β-酪蛋白掺入了去磷酸化的HeLa细胞提取物中。复合材料固有的亲水性使它具有更高的选择性，高达2000倍的复杂度，对磷肽的灵敏度低至1个飞摩尔。除了将金属离子固定在IMAC复合材料上之外，还测试了复合材料中基础材料（即纤维素和二氧化硅）的作用，以帮助其优化最佳样品制备方案。随着连续洗脱条件的引入，使用单批提取方法从蛋黄消化液中富集并鉴定了磷脂和磷酸肽。使用MALDI-MS分析磷酸肽，而使用新设计的金纳米颗粒和碳基材料通过LDI-MS分析低分子量磷脂。磷酸肽也从复杂的血清消化物中富集，并与前列腺癌有关。作为反相（RP）材料，它提供了亲水性复合材料和疏水性C18链的组合。与商业材料相比，高盐浓度的有效脱盐，改进的亲水性肽回收率和高序列覆盖率使纤维素-二氧化硅C18成为内部经济合成的更好的脱盐工具。

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