A Facile Hydrophilic Coating to Construct Immobilized Metal Affinity Chromatography for Protein Adsorption

IF 2.8 3区工程技术 Q2 CHEMISTRY, ANALYTICAL

Journal of separation science Pub Date : 2025-07-12 DOI:10.1002/jssc.70219

Xiao-Bei Huang, Yong-Ming Wei, Lin Yu, Zhen-Liang Xu, Hu Yang

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Abstract

Immobilized metal affinity chromatography is a green method applied to the separation and purification of proteins and biomolecules in downstream processing. With its unique internal pores, the membrane carriers have more active sites than typical magnetic nanoparticles and beads. It also greatly overcomes the mass-transfer resistance brought by traditional column chromatography. In this study, polysulfone was selected as the pristine membrane. Tannic acid and 3-aminopropyltriethoxysilane coatings were employed to construct the secondary reaction platform, which was then covalently coupled with polyethyleneimine and grafted with iminodiacetic acid groups to chelate Cu²⁺. Bovine serum albumin was utilized as the adsorption protein to adjust and optimize the modification parameters. The results demonstrated that the adsorption amount per unit membrane area reached 0.19 mg/cm² under the solution condition of pH 8.5. The adsorption thermodynamics followed the Langmuir model, and the corresponding model parameters were obtained.

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构建固定化金属亲和层析用于蛋白质吸附的易溶亲水涂层

固定化金属亲和色谱法是一种绿色的方法，应用于蛋白质和生物分子的分离纯化。由于其独特的内部孔隙，膜载体比典型的磁性纳米颗粒和磁珠具有更多的活性位点。它还极大地克服了传统柱色谱法带来的传质阻力。本研究选择聚砜作为原始膜。采用单宁酸和3-氨基丙基三乙氧基硅烷涂层构建二级反应平台，与聚乙烯亚胺共价偶联，接枝亚胺二乙酸基，螯合Cu2+。以牛血清白蛋白为吸附蛋白，对修饰参数进行调整和优化。结果表明，在pH为8.5的溶液条件下，单位膜面积的吸附量达到0.19 mg/cm2。吸附热力学遵循Langmuir模型，得到了相应的模型参数。

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来源期刊

Journal of separation science 化学-分析化学

CiteScore

6.30

自引率

16.10%

发文量

408

审稿时长

1.8 months

期刊介绍： The Journal of Separation Science (JSS) is the most comprehensive source in separation science, since it covers all areas of chromatographic and electrophoretic separation methods in theory and practice, both in the analytical and in the preparative mode, solid phase extraction, sample preparation, and related techniques. Manuscripts on methodological or instrumental developments, including detection aspects, in particular mass spectrometry, as well as on innovative applications will also be published. Manuscripts on hyphenation, automation, and miniaturization are particularly welcome. Pre- and post-separation facets of a total analysis may be covered as well as the underlying logic of the development or application of a method.