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具有为蛋白质和质粒DNA分离而设计的形态的电纺离子交换吸附剂的制备。

Fabrication of electrospun ion exchanger adsorbents with morphologies designed for the separation of proteins and plasmid DNA.

作者信息

Ovari Gyorgy, Johnson Thomas F, Foroutan Farzad, Malmquist Gunnar, Townsend Matthew, Bracewell Daniel G

机构信息

Department of Biochemical Engineering, University College London, Gower Street, London WC1E 6BT UK.

Department of Biochemical Engineering, University College London, Gower Street, London WC1E 6BT UK.

出版信息

J Chromatogr A. 2024 Oct 11;1734:465268. doi: 10.1016/j.chroma.2024.465268. Epub 2024 Aug 17.

Abstract

Electrospun cellulose adsorbents are an emergent class of materials applied to a variety of bioprocess separations as an analogue to conventional packed bed chromatography. Electrospun adsorbents have proven to be effective as rapid cycling media, enabling high throughput separation of proteins and viral vectors without compromising selectivity and recovery. However, there is a current lack of knowledge in relation to the manipulation and control of electrospun adsorbent structure with function and performance to cater to the separation needs of emerging, diverse biological products. In this study, a series of electrospun cellulose adsorbents were fabricated by adjusting their manufacturing conditions. A range of fiber diameters (400 to 600 nm) was created by changing the electrospinning polymer solution. Additionally, a range of porosities (0.4 to 0.7 v/v) was achieved by varying the laminating pressures on the electrospun sheets. The adsorbents were functionalized with different degrees of quaternary amine ligand density to create 18 prototype anion exchangers. Their morphology was characterized by BET nitrogen adsorption surface area, X-ray computed tomography, capillary flow porometry and scanning electron microscopy measurements. The physical characteristics of the adsorbents were used in an adapted semi-empirical model and compared to measured permeability data. Permeabilities of prototypes ranged from 10 to 10 mDarcy. The measured data showed good adherence to modelled data with possible improvements in acquiring wet adsorbent characteristics instead of dried material. Finally, the electrospun adsorbents were characterized for their binding capacity of model proteins of different sizes (diameters of 3.5 nm and 8.9 nm) and plasmid DNA. Static binding capacities ranged from 5 mg/ml to 25 mg/ml for the proteins and plasmid DNA and showed <20 % deviation from monolayer coverage based on BET surface area. Therefore, it was concluded that the electrospun adsorbents most likely adsorb monolayers of proteins and plasmid DNA on the surface with minimal steric hindrance.

摘要

电纺纤维素吸附剂是一类新兴材料,作为传统填充床色谱的类似物应用于各种生物过程分离。电纺吸附剂已被证明是有效的快速循环介质,能够在不影响选择性和回收率的情况下实现蛋白质和病毒载体的高通量分离。然而,目前在通过功能和性能来操纵和控制电纺吸附剂结构以满足新兴的、多样化生物产品的分离需求方面缺乏相关知识。在本研究中,通过调整其制造条件制备了一系列电纺纤维素吸附剂。通过改变电纺聚合物溶液,产生了一系列纤维直径(400至600纳米)。此外,通过改变电纺片材上的层压压力,实现了一系列孔隙率(0.4至0.7 v/v)。这些吸附剂用不同程度的季胺配体密度进行功能化,以创建18种原型阴离子交换剂。通过BET氮吸附表面积、X射线计算机断层扫描、毛细管流动孔隙率测定和扫描电子显微镜测量对其形态进行了表征。吸附剂的物理特性被用于一个改进的半经验模型,并与测量的渗透率数据进行比较。原型的渗透率范围为10至10毫达西。测量数据显示与模型数据具有良好的一致性,在获取湿吸附剂特性而非干燥材料方面可能还有改进空间。最后,对电纺吸附剂针对不同大小(直径分别为3.5纳米和8.9纳米)的模型蛋白质和质粒DNA的结合能力进行了表征。蛋白质和质粒DNA的静态结合容量范围为5毫克/毫升至25毫克/毫升,基于BET表面积,与单层覆盖的偏差小于20%。因此,得出的结论是,电纺吸附剂很可能在表面以最小的空间位阻吸附蛋白质和质粒DNA的单层。

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