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Implications from the grafting density and ionic capacity effects on protein adsorption to poly (N,N-dimethylaminopropyl acrylamide)-grafted Sepharose FF
Biochemical Engineering Journal ( IF 3.9 ) Pub Date : 2020-04-01 , DOI: 10.1016/j.bej.2020.107546
Changsen Li , Xianxiu Li , Yang Liu , Yan Sun

Abstract Our previous study reported a novel protein ion-exchanger, N,N-dimethylaminopropyl acrylamide (DMAPAA)-grafted Sepharose FF, of very high protein adsorption capacity and uptake rate. To gain new insight into the polymer grafting effects on protein adsorption and chromatography, 18 DMAPAA-grafted anion exchangers of four grafting densities and various chain lengths (i.e., ionic capacities, ICs) were prepared and named as FF-Brx-pDMAPAAn (x represents grafting density and n stands for IC in mmol/L). It was found that an optimal IC existed at each grafting density to present a maximal capacity. Of the four optimal-IC resins, FF-Br3-pDMAPAA587 showed the highest capacity (377 mg/mL), but the protein uptake rate, defined as the ratio of effective pore diffusivity to free diffusivity (De/D0), was smaller than the resin of a similar ionic capacity but lower grafting density (FF-Br2-pDMAPAA592). Consequently, the dynamic binding capacity (DBC) of FF-Br2-pDMAPAA592 was higher than that of FF-Br3-pDMAPAA587 at flow velocities higher than 450 cm/h. Moreover, FF-Br3-pDMAPAA587 showed higher adsorption capacity than FF-Br2-pDMAPAA592 at 0−150 mmol/L NaCl, leading to its higher DBC values as compared to FF-Br2-pDMAPAA592 in the same salt concentration range. Taken together, DMAPAA-grafted resins of different grafting densities would be useful for purifying feedstocks of different ionic strengths.

中文翻译:

接枝密度和离子容量对聚(N,N-二甲氨基丙基丙烯酰胺)接枝的 Sepharose FF 吸附蛋白质的影响

摘要 我们之前的研究报道了一种新型蛋白质离子交换剂 N,N-二甲基氨基丙基丙烯酰胺 (DMAPAA) 接枝的 Sepharose FF,具有非常高的蛋白质吸附能力和摄取率。为了深入了解聚合物接枝对蛋白质吸附和色谱的影响,制备了 18 种具有四种接枝密度和不同链长(即离子容量、IC)的 DMAPAA 接枝阴离子交换剂,并将其命名为 FF-Brx-pDMAPAAn(x 代表接枝密度,n 代表 IC,单位为 mmol/L)。发现在每个接枝密度下都存在最佳 IC 以呈现最大容量。在四种最佳 IC 树脂中,FF-Br3-pDMAPAA587 表现出最高的容量 (377 mg/mL),但蛋白质吸收率,定义为有效孔扩散率与自由扩散率的比率 (De/D0),小于具有相似离子容量但接枝密度较低的树脂 (FF-Br2-pDMAPAA592)。因此,在流速高于 450 cm/h 时,FF-Br2-pDMAPAA592 的动态结合容量 (DBC) 高于 FF-Br3-pDMAPAA587。此外,FF-Br3-pDMAPAA587 在 0-150 mmol/L NaCl 下显示出比 FF-Br2-pDMAPAA592 更高的吸附容量,导致在相同盐浓度范围内与 FF-Br2-pDMAPAA592 相比其 DBC 值更高。总之,不同接枝密度的 DMAPAA 接枝树脂可用于纯化不同离子强度的原料。FF-Br3-pDMAPAA587 在 0-150 mmol/L NaCl 下显示出比 FF-Br2-pDMAPAA592 更高的吸附容量,导致在相同盐浓度范围内与 FF-Br2-pDMAPAA592 相比其 DBC 值更高。总之,不同接枝密度的 DMAPAA 接枝树脂可用于纯化不同离子强度的原料。FF-Br3-pDMAPAA587 在 0-150 mmol/L NaCl 下显示出比 FF-Br2-pDMAPAA592 更高的吸附容量,导致在相同盐浓度范围内与 FF-Br2-pDMAPAA592 相比其 DBC 值更高。总之,不同接枝密度的 DMAPAA 接枝树脂可用于纯化不同离子强度的原料。
更新日期:2020-04-01
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