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采用磁性介孔稀土硅酸盐微球对牛血红蛋白进行选择性分离。

Selective separation of bovine hemoglobin using magnetic mesoporous rare-earth silicate microspheres.

机构信息

MOE Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology, Ministry of Education, Department of Chemistry, Tsinghua University, Beijing, 100084, China.

MOE Key Laboratory of Bioorganic Phosphorus Chemistry and Chemical Biology, Ministry of Education, Department of Chemistry, Tsinghua University, Beijing, 100084, China.

出版信息

Talanta. 2019 Nov 1;204:792-801. doi: 10.1016/j.talanta.2019.06.069. Epub 2019 Jun 19.

Abstract

Selective separation of heme proteins and peptides from complicated biological samples before comprehensive identification and characterization of intact biomolecules which are low stoichiometry is indispensable for ongoing proteomics. Here, three magnetic mesoporous rare-earth silicate (rare-earth = Er, Tm, Yb) microspheres prepared by facile solvothermal method were used as novel adsorbents for the selective isolation of bovine hemoglobin (BHb), and magnetic mesoporous ytterbium silicate showed apparent adsorption efficiency in isolating BHb with a adsorption capacity of 304.4 mg/g after the comparison. The retained BHb could be eluted by using NaCO as stripping reagent. Meanwhile, Circular dichroism spectra illustrated that the microspheres posed no effect on the secondary structure of BHb. The resultant magnetic particles were characterized by transmission electron microscope, scanning electron microscope, x-ray powder diffraction, vibrating sample magnetometer, Fourier transform infrared spectroscopy, N adsorption-desorption isotherm and zeta potential. The as-prepared magnetic microspheres showed high specificity for the separation of BHb from bovine plasma as corroborated by SDS-PAGE and MALDI-TOF MS analysis, which would be expected to possess potential application in isolation of other heme proteins in complex biological samples.

摘要

在对低丰度的完整生物分子进行全面鉴定和表征之前,从复杂的生物样品中选择性地分离血红素蛋白和肽对于正在进行的蛋白质组学研究是必不可少的。在这里,通过简便的溶剂热法制备了三种磁性介孔稀土硅酸盐(稀土= Er、Tm、Yb)微球,用作新型吸附剂,用于选择性分离牛血红蛋白(BHb),经过比较,磁性介孔 ytterbium 硅酸盐显示出明显的吸附效率,吸附容量为 304.4 mg/g。保留的 BHb 可以用 NaCO 作为洗脱试剂洗脱。同时,圆二色光谱表明微球对 BHb 的二级结构没有影响。通过透射电子显微镜、扫描电子显微镜、X 射线粉末衍射、振动样品磁强计、傅里叶变换红外光谱、N 吸附-脱附等温线和 zeta 电位对所得磁性颗粒进行了表征。SDS-PAGE 和 MALDI-TOF MS 分析证实,所制备的磁性微球对从牛血浆中分离 BHb 具有高特异性,有望在复杂生物样品中分离其他血红素蛋白方面具有潜在应用。

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