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.
Anal Chim Acta. 2017 Feb 15;954:129-139. doi: 10.1016/j.aca.2016.11.065. Epub 2016 Dec 3.
A weak cation exchange liquid chromatography stationary phase (nylon-COOH) was prepared by grafting polyacrylic acid on to native nylon 6 capillary-channeled polymer (C-CP) fibers via a microwave-assisted radical polymerization. To the best of our knowledge, this is the first study of applying microwave-assisted grafting polymerization to affect nylon material for protein separation. The C-CP fiber surfaces were characterized by attenuated total reflection (ATR) infrared spectroscopy and scanning electron microscope (SEM). The anticipated carbonyl peak at 1722.9 cm was found on the nylon-COOH fibers, but was not found on the native fiber, indicating the presence of the polyacrylic acid on nylon fibers after grafting. The nylon-COOH phase showed a ∼12× increase in lysozyme dynamic binding capacity (∼12 mg mL) when compared to the native fiber phase (∼1 mg mL). The loading capacity of the nylon-COOH phase is nearly independent of the lysozyme loading concentration (0.05-1 mg mL) and the mobile phase linear velocity (7.3-73 mm s). The reproducibility of the lysozyme recovery from the nylon-COOH (RSD = 0.3%, n = 10) and the batch-to-batch variability in the functionalization (RSD = 3%, n = 5) were also investigated, revealing very high levels of consistency. Fast baseline separations of myoglobin, α-chymotrypsinogen A, cytochrome c and lysozyme were achieved using the nylon-COOH column. It was found that a 5× increase in the mobile phase linear velocity (7.3-to-36.5 mm s) had little effect on the separation resolution. The microwave-assisted grafting polymerization has great potential as a generalized surface modification methodology across the applications of C-CP fibers.
一种弱阳离子交换的液相色谱固定相（尼龙-COOH）是通过微波辅助自由基聚合将聚丙烯酸接枝到天然尼龙 6 毛细管通道聚合物（C-CP）纤维上而制备的。据我们所知，这是首次将微波辅助接枝聚合应用于影响蛋白质分离用尼龙材料的研究。C-CP 纤维表面的特性通过衰减全反射（ATR）红外光谱和扫描电子显微镜（SEM）进行了表征。在尼龙-COOH 纤维上发现了预期的 1722.9 cm 处的羰基峰，但在天然纤维上未发现，表明接枝后尼龙纤维上存在聚丙烯酸。与天然纤维相比，尼龙-COOH 相的溶菌酶动态结合容量（12 mg mL）增加了约 12 倍（1 mg mL）。尼龙-COOH 相的负载容量几乎与溶菌酶的负载浓度（0.05-1 mg mL）和流动相线性速度（7.3-73 mm s）无关。还研究了从尼龙-COOH 中回收溶菌酶的重现性（RSD = 0.3%，n = 10）和官能化的批间可变性（RSD = 3%，n = 5），结果表明一致性非常高。使用尼龙-COOH 柱可实现肌红蛋白、α-糜蛋白酶原 A、细胞色素 c 和溶菌酶的快速基线分离。结果发现，流动相线性速度增加 5 倍（从 7.3 增加到 36.5 mm s）对分离分辨率几乎没有影响。微波辅助接枝聚合作为一种通用的表面改性方法，在 C-CP 纤维的应用中具有巨大的潜力。
