Research Center for Analytical Sciences, College of Sciences, Northeastern University, Box 332, Shenyang 110819, China.
Research Center for Analytical Sciences, College of Sciences, Northeastern University, Box 332, Shenyang 110819, China.
Talanta. 2016 Oct 1;159:23-28. doi: 10.1016/j.talanta.2016.06.005. Epub 2016 Jun 3.
A novel polyoxometalate (POM)-based hybrid P8W48-APTS is prepared by the solidification of super-lacunary P8W48O184(40-) polyoxoanions with APTS in an acidic medium. The oxygen (O(-)) atoms in P8W48O184(40-) are bound to silicon atoms in APTS by the formation of Si-O linkage through dehydration condensation. The solidification is confirmed by characterizations with XRD, FT-IR, TGA, SEM and EDXS. Selective isolation of proteins of interest, hemoglobin (Hb) in this case, from complex sample matrices is achieved by using P8W48-APTS hybrid as adsorbent under controlled conditions. 5.0mg of P8W48-APTS hybrid results in an adsorption efficiency of 93% for 100mgL(-1) hemoglobin in 1.0mL sample solution at pH 7. The adsorption behavior of Hb onto P8W48-APTS hybrid fits Langmuir adsorption model, corresponding to an adsorption capacity of 355.0mgg(-1). The retained Hb could be readily recovered with either a SDS solution (0.1molL(-1)) or a Na3PO4 (0.1molL(-1)) solution as stripping reagent, providing recoveries of 94.6% or 83.9%, respectively. The biological activity of Hb remains 96.7% after an adsorption/desorption process (with elution by SDS), which illustrates virtually no change on the conformation of hemoglobin. The P8W48-APTS hybrid has been applied for the selective adsorption of Hb from human whole blood, and the results are demonstrated by SDS-PAGE assay.
一种新型的多金属氧酸盐(POM)基杂化 P8W48-APTS 是通过在酸性介质中使超笼 P8W48O184(40-)多氧阴离子与 APTS 固化而制备的。P8W48O184(40-)中的氧(O(-))原子通过脱水缩合与 APTS 中的硅原子形成 Si-O 键连接。通过 XRD、FT-IR、TGA、SEM 和 EDXS 的特征证实了固化。在控制条件下,将 P8W48-APTS 杂化作为吸附剂,从复杂的样品基质中选择性分离出感兴趣的蛋白质,在这种情况下是血红蛋白(Hb)。在 pH 7 下,5.0mg 的 P8W48-APTS 杂化可使 1.0mL 样品溶液中 100mgL(-1)的血红蛋白的吸附效率达到 93%。Hb 吸附到 P8W48-APTS 杂化上的吸附行为符合 Langmuir 吸附模型,对应于 355.0mgg(-1)的吸附容量。保留的 Hb 可以很容易地用 SDS 溶液(0.1molL(-1))或 Na3PO4(0.1molL(-1))溶液作为洗脱试剂回收,回收率分别为 94.6%或 83.9%。Hb 的生物活性在吸附/解吸过程(用 SDS 洗脱)后保持 96.7%,这表明血红蛋白的构象几乎没有变化。P8W48-APTS 杂化已应用于从人全血中选择性吸附 Hb,并通过 SDS-PAGE 分析进行了验证。