Electrospun polyethersulfone affinity membrane: Membrane preparation and performance evaluation

被引:59
作者
Ma, Zuwei [1 ]
Lan, Zhengwei [1 ]
Matsuura, Takeshi [2 ]
Ramakrishna, Seeram [1 ]
机构
[1] Natl Univ Singapore, Nanosci & Nanotechnol Initiat, Singapore 117576, Singapore
[2] Univ Ottawa, Dept Chem Engn, Ottawa, ON K1N 6N5, Canada
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2009年 / 877卷 / 29期
关键词
Affinity membrane; Frontal analysis; Breakthrough; Electrospinning; Polyethersulphone (PES); IgG purification; Spin column; Protein A/C; Plate model; CELLULOSE NANOFIBER; FRONTAL ANALYSIS; PROTEIN-A/G; PURIFICATION; IMMOBILIZATION; CHROMATOGRAPHY; ADSORPTION; ALBUMIN; ACID;
D O I
10.1016/j.jchromb.2009.09.019
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Non-woven polyethersulfone (PES) membranes were prepared by electrospinning. After heat treatment and surface activation, the membranes were covalently functionalized with ligands to be used as affinity membranes. The membranes were characterized in terms of fiber diameter, porosity, specific area, pore size, ligand density and binding capacities. To evaluate the binding efficiency of the membrane, dynamic adsorption of bovine serum albumin (BSA) on the Cibacron blue F3GA (CB) functionalized PES membrane was studied. Experimental breakthrough curves were fitted with the theoretical curves based on the plate model to estimate plate height (Hp) of the affinity membrane. The high value of H(p) (1.6-8 cm) of the affinity membrane implied a poor dynamic binding efficiency, which can be explained by the intrinsic microstructures of the material. Although the electrospun membrane might not be an ideal candidate for the preparative affinity membrane chromatography for large-scale production, it still can be used for fast small-scale protein purification in which a highly efficient binding is not required. Spin columns packed with protein A/G immobilized PES membranes were demonstrated to be capable of binding IgG specifically. SDS-PAGE results demonstrated that the PES affinity membrane had high specific binding selectivity for IgG molecules and low non-specific protein adsorption. Compared with other reported affinity membranes, the PES affinity membrane had a comparable IgG binding capacity of 4.5 mg/ml, and had a lower flow through pressure drop due to its larger pore size. In conclusion, the novel PES affinity membrane is an ideal spin Column packing material for fast protein purification. (C) 2009 Elsevier B.V. All rights reserved.
引用
收藏
页码:3686 / 3694
页数:9
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