CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, P. R. China.
University of Chinese Academy of Sciences, Beijing, P. R. China.
J Sep Sci. 2019 Feb;42(3):744-753. doi: 10.1002/jssc.201800969. Epub 2018 Dec 13.
Employing immobilized metal-ion affinity chromatography and magnetic separation could ideally provide a useful analytical strategy for purifying His-tagged protein. In the current study, a facile route was designed to prepare CMPEI-Ni @SiO @Fe O (CMPEI=carboxymethylated polyethyleneimine) magnetic nanoparticles composed of a strong magnetic core of Fe O and a Ni -immobilized carboxymethylated polyethyleneimine coated outside shell, which was formed by electrostatic interactions between polyanionic electrolyte of carboxymethylated polyethyleneimine and positively charged surface of 3-(trimethoxysilyl)propylamin modified SiO @Fe O . The resulting CMPEI-Ni @SiO @Fe O composite nanoparticles displayed well-uniform structure and high magnetic responsiveness. Hexa His-tagged peptides and purified His-tagged recombinant retinoid X receptor alpha were chosen as the model samples to evaluate the adsorption, capacity, and reusability of the composite nanoparticles. The results demonstrated the CMPEI-Ni @SiO @Fe O nanoparticles possessed rapid adsorption, large capacity, and good recyclability. The obtained nanoparticles were further used to purify His-tagged protein in practical environment. It was found that the nanoparticles could selectively capture His-tagged recombinant retinoid X receptor protein from complex cell lysate. Owing to its easy synthesis, large binding capacity, and good reusability, the prepared CMPEI-Ni @SiO @Fe O magnetic nanoparticles have great potential for application in biotechnological fields.
采用固定化金属离子亲和层析和磁分离技术，理想情况下可以为纯化 His 标签蛋白提供一种有用的分析策略。在本研究中，设计了一种简便的方法来制备 CMPEI-Ni@SiO@Fe3O4（CMPEI=羧甲基化聚乙烯亚胺）磁性纳米粒子，由 Fe3O4 强磁核和 Ni 固定化的羧甲基化聚乙烯亚胺涂覆的外壳组成，这是通过带负电荷的羧甲基化聚乙烯亚胺电解质与 3-（三甲氧基硅基）丙基氨基改性的 SiO@Fe3O4 的正电荷表面之间的静电相互作用形成的。所得的 CMPEI-Ni@SiO@Fe3O4 复合纳米粒子显示出均匀的结构和高的磁响应性。六 His 肽和纯化的 His 标签重组视黄醇 X 受体 α 被选为模型样品，以评估复合纳米粒子的吸附、容量和可重复使用性。结果表明，CMPEI-Ni@SiO@Fe3O4 纳米粒子具有快速吸附、大容量和良好的可重复使用性。所得纳米粒子进一步用于实际环境中 His 标签蛋白的纯化。结果发现，纳米粒子可以从复杂的细胞裂解物中选择性地捕获 His 标签重组视黄醇 X 受体蛋白。由于其合成简单、结合容量大、可重复使用性好，制备的 CMPEI-Ni@SiO@Fe3O4 磁性纳米粒子在生物技术领域具有很大的应用潜力。
