Department of Clinical Laboratory Medicine, Shanghai Tenth People's Hospital, School of Medicine, Tongji University, Shanghai, 200072, China.
Department of Clinical Laboratory Medicine, Shanghai Children's Medical Center, School of Medicine, Shanghai Jiaotong University, Shanghai, 200127, China.
Anal Bioanal Chem. 2021 May;413(11):2893-2901. doi: 10.1007/s00216-021-03215-9. Epub 2021 Mar 11.
Selective separation and enrichment of phosphoproteins possess the distinct clinical and biological importance in the diagnosis, treatment, and management of several fatal human diseases. In this study, a facile synthesis of titanium(IV) ion-immobilized arsenate-modified poly(glycidyl methacrylate) microparticles (denoted as Ti-arsenate-PGMA-MPs) was developed for the efficient enrichment of intact phosphoproteins found in biologically complex protein samples. By virtue of the strong interaction between the titanium ions immobilized on the surface of Ti-arsenate-PGMA-MPs and phosphate groups of phosphoproteins, Ti-arsenate-PGMA-MPs had a high saturated adsorption capacity for phosphoproteins (901 mg/g for β-casein), which was much higher than that of non-phosphoproteins (73.5 mg/g for BSA). Ti-arsenate-PGMA-MPs were characterized by SEM, TEM, and FT-IR, and the average particle diameter was about 2.5 μm with good dispersibility. Besides, the application of Ti-arsenate-PGMA-MPs in real biological samples was investigated by SDS-PAGE analysis, and the results showed that Ti-arsenate-PGMA-MPs were able to enrich phosphoproteins efficiently.
选择性分离和富集磷酸化蛋白质在几种致命人类疾病的诊断、治疗和管理中具有重要的临床和生物学意义。在本研究中，开发了一种简便的钛(IV)离子固定的砷酸盐修饰的聚(甲基丙烯酸缩水甘油酯)微球（表示为 Ti-arsenate-PGMA-MPs）的合成方法，用于从生物复杂蛋白质样品中有效地富集完整的磷酸化蛋白质。由于 Ti-arsenate-PGMA-MPs 表面固定的钛离子与磷酸化蛋白质的磷酸基团之间的强相互作用，Ti-arsenate-PGMA-MPs 对磷酸化蛋白质具有高的饱和吸附容量（β-酪蛋白为 901mg/g），远高于非磷酸化蛋白质（BSA 为 73.5mg/g）。Ti-arsenate-PGMA-MPs 通过 SEM、TEM 和 FT-IR 进行了表征，平均粒径约为 2.5μm，具有良好的分散性。此外，还通过 SDS-PAGE 分析研究了 Ti-arsenate-PGMA-MPs 在实际生物样品中的应用，结果表明 Ti-arsenate-PGMA-MPs 能够有效地富集磷酸化蛋白质。
