Abstract

Hydrophobic interaction chromatography (HIC) as an indispensable method for protein purification has attracted considerable attentions of researchers as well as biopharmaceutical industries. However, the low binding capacity and slow adsorption rate of the currently available HIC media lead to a little supply and high price of the highly purified proteins. Herein, nanofibrous membranes with hydrophobic binding sites were developed for HIC by directly coupling phenyl glycidyl ether on the hydrolyzed cellulose acetate nanofiber membrane (cellulose-phenyl NFM). Scanning electron microscope (SEM), water contact angle (WCA), Fourier transform infrared (FTIR), thermogravimetric analysis (TGA), Brunauer–Emmett–Teller (BET) surface area analysis and capillary flow porometer (CFP) were applied to evaluate the physically and chemically structural transformation. The obtained cellulose-phenyl NFMs showed a proper hydrophilcity (WCA = 37°), a relatively high BET surface area (3.6 times the surface area of commercial fibrous membranes), and tortuous-channel structure with through-hole size in the range of 0.25–1.2 μm, which led to a little non-specificity adsorption, high bovine serum albumin adsorption capacity of 118 mg g⁻¹, fast adsorption process within 12 h, good long-term stability and reusability. Moreover, compared with traditional modification methods which always include activation and graft two steps, direct coupling method is more efficient for HIC media fabrication. Therefore, cellulose-phenyl NFMs with outstanding protein adsorption performance could be a kind of promising candidate for HIC.

Graphic Abstract

中文翻译：

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苯基缩水甘油醚修饰的纤维素纳米纤维膜对牛血清白蛋白的高效吸附

摘要

疏水相互作用色谱法（HIC）作为蛋白质纯化必不可少的方法已经引起了研究人员以及生物制药行业的广泛关注。然而，目前可用的HIC介质的低结合容量和缓慢的吸附速率导致高度纯化的蛋白质供应少且价格高。本文中，通过将苯基缩水甘油醚直接偶联在水解的乙酸纤维素纳米纤维膜（纤维素-苯基NFM）上，开发了具有疏水性结合位点的纳米纤维膜用于HIC。扫描电子显微镜（SEM），水接触角（WCA），傅立叶变换红外（FTIR），热重分析（TGA），Brunauer-Emmett-Teller（BET）表面积分析和毛细管流动孔度计（CFP）用于评估物理和化学结构的转变。所得的纤维素-苯基NFM具有适当的亲水性（WCA = 37°），相对较高的BET表面积（3.6倍于商用纤维膜的表面积）和曲折通道结构，通孔尺寸在0.25范围内–1.2μm，导致少量非特异性吸附，高牛血清白蛋白吸附能力为118 mg g^-1，在12小时内快速吸附，长期稳定性和可重复使用性良好。而且，与始终包括激活和接枝两个步骤的传统修饰方法相比，直接耦合方法对于HIC介质的制造更为有效。因此，具有优异蛋白质吸附性能的纤维素-苯基NFMs可能是HIC的一种有希望的候选物。

图形摘要