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表面功能化的静电纺丝碳纳米纤维垫作为一种新型的蛋白质吸附/纯化介质,具有高容量和高通量的特点。

Surface-functionalized electrospun carbon nanofiber mats as an innovative type of protein adsorption/purification medium with high capacity and high throughput.

机构信息

Department of Chemical and Biological Engineering, South Dakota School of Mines and Technology, Rapid City, SD 57701, USA.

出版信息

J Chromatogr A. 2011 Dec 16;1218(50):8989-95. doi: 10.1016/j.chroma.2011.10.024. Epub 2011 Oct 15.

DOI:10.1016/j.chroma.2011.10.024
PMID:22047820
Abstract

Due to recent advances in the production of biotherapeutics, high capacity, high throughput adsorption media for efficient and economic separation of these medically important products are in great demand. One option that has been evaluated extensively is membrane/mat adsorption. While these media allow for rapid adsorption (due to the decreased internal diffusion) and high throughput processing (due to the open porous structure), they often suffer from low capacity and poor enrichment factors. Herein, we report the fabrication, characterization, and protein adsorption evaluation of an innovative type of membrane/mat adsorption media based on electrospun carbon nanofibers. By surface-functionalization of these nanofibers with a weak acid cation-exchange ligand, the capacity was doubled for binding a model protein (i.e., lysozyme) compared to commercial products; and the capacity value was over 200 mg lysozyme per gram of adsorption media. Meanwhile, the thin nanofibers (having diameters of ~300 nm) along with open pores among nanofibers in the mats (having sizes of ~10-15 μm) allowed for higher operating flow rates and lower pressure drops. Furthermore, the incorporation of higher ligand density and the addition of a non-ionic surfactant (i.e., Triton X-305) into the adsorption buffer eliminated the non-specific binding of a competing protein (bovine serum albumin). In combination, this study suggested that electrospun carbon nanofiber adsorption media would provide a promising alternative to packed resin beds for bioseparations.

摘要

由于生物治疗药物生产的最新进展,对高效经济分离这些医学重要产品的大容量、高通量吸附介质的需求很大。已经广泛评估的一种选择是膜/垫吸附。虽然这些介质允许快速吸附(由于内部扩散减少)和高通量处理(由于开放多孔结构),但它们通常容量低且富集因子差。在此,我们报告了一种基于静电纺丝碳纤维的新型膜/垫吸附介质的制造、表征和蛋白质吸附评价。通过将这些纳米纤维用弱酸阳离子交换配体进行表面功能化,与商业产品相比,结合模型蛋白(即溶菌酶)的容量增加了一倍;吸附介质每克的容量值超过 200 毫克溶菌酶。同时,薄纳米纤维(直径约 300nm)以及垫子中纳米纤维之间的开放孔(尺寸约为 10-15μm)允许更高的操作流速和更低的压降。此外,在吸附缓冲液中加入更高的配体密度和非离子表面活性剂(即 Triton X-305)消除了竞争蛋白(牛血清白蛋白)的非特异性结合。总之,这项研究表明,静电纺丝碳纤维吸附介质将为生物分离提供一种有前途的替代填充树脂床的方法。

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