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用于蛋白质阴离子交换色谱的聚乙烯亚胺改性聚对苯二甲酸乙二酯毛细管通道聚合物纤维

Polyethylenimine modified poly(ethylene terephthalate) capillary channeled-polymer fibers for anion exchange chromatography of proteins.

作者信息

Jiang Liuwei, Jin Yi, Marcus R Kenneth

机构信息

Clemson University, Department of Chemistry, Biosystems Research Complex, Clemson, SC 29634, United States.

Clemson University, Department of Chemistry, Biosystems Research Complex, Clemson, SC 29634, United States.

出版信息

J Chromatogr A. 2015 Sep 4;1410:200-9. doi: 10.1016/j.chroma.2015.07.102. Epub 2015 Jul 29.

Abstract

Native poly(ethylene terephthalate) (PET) capillary-channeled polymer (C-CP) fibers have been previously studied as stationary phases for reversed phase and affinity protein separations. In this study, surface modified PET C-CP fibers were evaluated for the anion exchange separation of proteins. The native PET C-CP fibers were aminated using polyethylenimine (PEI) followed by a 1,4-butanediol diglycidyl ether (BUDGE) cross-linking step. Subsequent PEI/BUDGE treatments can be employed to further develop the polyamine layer on the fiber surfaces. The PEI densities of the modified fibers were quantified through the ninhydrin reaction, yielding values of 0.43-0.89μmolg(-1). The surface modification impact on column permeability was found to be 0.66×10(-11) to 1.33×10(-11)m(2), depending on the modification time and conditions. The dynamic binding capacities of the modified fiber media were determined to be 1.99-8.54mgmL(-1) bed volume, at linear velocities of 88-438cmmin(-1) using bovine serum albumin as the model protein. It was found that increasing the mobile phase linear velocity (up to 438cmmin(-1)) had no effect on the separation quality for a synthetic protein mixture, reflecting the lack of van Deemter C-term effects for the C-CP fiber phase. The low-cost, easy modification method and the capability of fast protein separation illustrate great potential in the use of PEI/BUDGE-modified PET C-CP fibers for high-throughput protein separation and downstream processing.

摘要

天然聚对苯二甲酸乙二酯(PET)毛细管通道聚合物(C-CP)纤维此前已被研究用作反相和亲和蛋白质分离的固定相。在本研究中,对表面改性的PET C-CP纤维进行了蛋白质阴离子交换分离评估。天然PET C-CP纤维先用聚乙烯亚胺(PEI)胺化,然后进行1,4-丁二醇二缩水甘油醚(BUDGE)交联步骤。随后可采用PEI/BUDGE处理进一步在纤维表面形成多胺层。通过茚三酮反应对改性纤维的PEI密度进行了定量,得到的值为0.43 - 0.89μmol g⁻¹。发现表面改性对柱渗透率的影响为0.66×10⁻¹¹至1.33×10⁻¹¹ m²,这取决于改性时间和条件。以牛血清白蛋白为模型蛋白,在88 - 438 cm min⁻¹的线性流速下,测定改性纤维介质的动态结合容量为1.99 - 8.54 mg mL⁻¹床体积。结果发现,增加流动相线性流速(高达438 cm min⁻¹)对合成蛋白质混合物的分离质量没有影响,这表明C-CP纤维相对范德姆特C项效应不敏感。低成本、易于改性的方法以及快速蛋白质分离的能力表明,PEI/BUDGE改性的PET C-CP纤维在高通量蛋白质分离和下游处理方面具有巨大潜力。

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