Farzi-Khajeh Hamed, Safa Kazem D, Dastmalchi Siavoush
Organosilicon Research Laboratory, Faculty of Chemistry, University of Tabriz, Tabriz, Iran.
Organosilicon Research Laboratory, Faculty of Chemistry, University of Tabriz, Tabriz, Iran.
J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Sep 1;1061-1062:26-33. doi: 10.1016/j.jchromb.2017.06.048. Epub 2017 Jun 29.
Recent studies of magnetic carrier technology have focused on its applications in separation and purification technologies, due to easy separation of the target from the reaction medium by applying an external magnetic field. In the present study, FeO superparamagnetic nanoparticles were prepared to utilize a chemical co-precipitation method, then the surfaces of the nanoparticles were modified with arsanilic acid derivatives which were used as the specific nanocarriers for the affinity purification of alkaline phosphatase from the hen's egg yolk. The six different types of magnetic nanocarriers with varied lengths of the linkers were obtained. All samples were characterized step by step and validated using FTIR, SEM, EDX, VSM and XRD analysis methods As the results were shown, the use of inflexible tags with long linkers on the surface of the nanocarrier could lead to better results for separation of alkaline phosphatase from the hen's egg yolk with 76.2% recovery and 1361.7-fold purification. The molecular weight of the purified alkaline phosphatase was estimated to be 68kDa by SDS-PAGE. The results of this study showed that the novel magnetic nanocarriers were capable of purifying alkaline phosphatase in a practically time and cost effective way.
最近对磁性载体技术的研究集中在其在分离和纯化技术中的应用上，这是因为通过施加外部磁场可以轻松地将目标物与反应介质分离。在本研究中，采用化学共沉淀法制备了FeO超顺磁性纳米颗粒，然后用对氨基苯胂酸衍生物对纳米颗粒表面进行修饰，这些衍生物用作从鸡蛋黄中亲和纯化碱性磷酸酶的特定纳米载体。获得了六种不同类型的具有不同长度连接子的磁性纳米载体。使用FTIR、SEM、EDX、VSM和XRD分析方法对所有样品进行了逐步表征和验证。结果表明，在纳米载体表面使用带有长连接子的刚性标签能够从鸡蛋黄中分离碱性磷酸酶，回收率为76.2%，纯化倍数为1361.7倍，效果更好。通过SDS-PAGE估计纯化后的碱性磷酸酶分子量为68kDa。本研究结果表明，新型磁性纳米载体能够以切实可行的方式在时间和成本上高效地纯化碱性磷酸酶。
