The work describes the preparation of a new magnetic phase for batch enrichment of phosphopeptides. The material exploits the advantages of magnetic solid phase extraction and couples them with the most employed approach for phosphopeptide enrichment, i.e. Ti⁴⁺-IMAC. In order to immobilize Ti⁴⁺ ions on the surface of the magnetite nanoparticles, they were first covered by a silica shell and then modified to expose at the surface bromine containing groups. Glycidyl methacrylate was subsequently polymerized from these groups using the “grafting from” approach by the activator regenerated by electron transfer–atom transfer radical polymerization (ARGET-ATRP) technique. Finally, the glycidyl groups were reacted with iminodiacetic acid to functionalize the material with moieties suitable for coordination. The prepared material was extensively characterized and subsequently tested for enrichment of a bovine serum albumin mixture with casein to ascertain its potential. With positive results, the new magnetic polymeric material was further employed to set up an enrichment method on yeast protein digest based on shotgun proteomics. The sample to phase ratio was optimized and the best condition compared to a commercial TiO₂ spin column. At the end of the comparison, the new material proved better and could enrich a larger total number of phosphopeptides with increased selectivity. All these conclusions and the test performed on a real complex sample within the final shotgun application further support the applicability of the new material in phosphopeptide analysis of real matrices.

这项工作描述了用于批量富集磷酸肽的新磁性相的制备。该材料利用了磁性固相萃取的优势，并将其与最常用的磷酸肽富集方法（即Ti ⁴⁺ -IMAC）结合在一起。为了固定Ti ⁴⁺磁铁矿纳米颗粒表面的离子，它们首先被二氧化硅壳覆盖，然后被修饰以在表面暴露出含溴基团。甲基丙烯酸缩水甘油酯随后通过“接枝”方法由通过电子转移-原子转移自由基聚合（ARGET-ATRP）技术再生的活化剂从这些基团聚合而来。最后，使缩水甘油基与亚氨基二乙酸反应，以使材料具有适合于配位的部分官能化。对所制备的材料进行了广泛的表征，随后测试了酪蛋白对牛血清白蛋白混合物的富集，以确定其潜力。取得了积极的成果，新的磁性高分子材料被进一步用于建立基于shot弹枪蛋白质组学的酵母蛋白质消化的富集方法。₂旋转柱。在比较结束时，新材料证明更好，并且可以通过增加选择性来富集更多总数的磷酸肽。所有这些结论和在最终shot弹枪应用程序中对真实复杂样品进行的测试进一步支持了新材料在真实基质的磷酸肽分析中的适用性。