反离子对部分电荷中和的聚乙烯亚胺接枝介质的蛋白质色谱行为的影响

doi:10.11949/j.issn.0438-1157.20170360

化工学报 ›› 2017, Vol. 68 ›› Issue (11): 4178-4185.DOI: 10.11949/j.issn.0438-1157.20170360

反离子对部分电荷中和的聚乙烯亚胺接枝介质的蛋白质色谱行为的影响

翟秋红, 余林玲, 孙彦

天津大学化工学院系统生物工程教育部重点实验室, 天津 300072

收稿日期:2017-04-10 修回日期:2017-07-11 出版日期:2017-11-05 发布日期:2017-11-05
通讯作者: 余林玲
基金资助:
国家自然科学基金项目（21406160，21236005，21621004）。

Effect of counterions on protein adsorption to partial neutralization poly(ethylenimine)-grafted Sepharose FF

ZHAI Qiuhong, YU Linling, SUN Yan

Department of Biochemical Engineering and Key Laboratory of Systems Bioengineering of Ministry of Education, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China

Received:2017-04-10 Revised:2017-07-11 Online:2017-11-05 Published:2017-11-05
Supported by:
supported by the National Natural Science Foundation of China (21406160, 21236005, 21621004).

摘要/Abstract

摘要：

前期研究表明部分电荷中和的聚乙烯亚胺接枝琼脂糖介质FF-PEI-R440具有较高静态吸附容量和快速传质速率。本文选取硫氰酸根离子、氯离子、磷酸氢根离子以及硫酸根离子这4种反离子，在离子强度为0.03、0.06 mol·L^-1下，研究其对FF-PEI-R440吸附与洗脱行为的影响。结果表明：在两种离子强度下，饱和吸附容量的增加顺序均为SCN^- < SO₄^2- < HPO₄^2- < Cl^-；在低离子强度下，除SCN^-外，反离子种类对传质速率和动态结合容量没有显著影响；在高离子强度下，传质速率的增加顺序为SCN^- < SO₄^2- ≈ Cl^- < HPO₄^2-，而动态结合容量的增加顺序与饱和吸附容量序列一致；两种离子强度下Cl^-均保持最高的动态结合容量；反离子种类对蛋白质洗脱行为没有显著影响。上述结果说明反离子主要影响FF-PEI-R440的吸附性能，而不影响洗脱，且流动相中选择Cl^-为反离子最利于FF-PEI-R440的实际柱色谱操作。

关键词: 离子交换色谱, 聚合物接枝, 反离子, 吸附平衡, 动力学, 动态结合容量, 线性梯度洗脱

Abstract:

The partially neutralized poly(ethylenimine)-grafted Sepharose FF, FF-PEI-R440, exhibited higher protein capacity and faster uptake rate than its starting resin FF-PEI-L740. In this work, the influence of counterions on protein adsorption onto and elution from the FF-PEI-R440 resin was investigated with sodium salts of SCN^-, Cl^-, HPO₄^2- and SO₄^2- at ionic strengths of 0.03 mol·L^-1 and 0.06 mol·L^-1. It was found that the static adsorption equilibrium of FF-PEI-R440 was significantly influenced by the counterions, and the adsorption capacity increased in the order of SCN^- < SO₄^2- < HPO₄^2- < Cl^- at the two ionic strengths. At the low ionic strength, the counterions had no significant effect on the uptake rate and dynamic binding capacity except for SCN^-. At high ionic strength, the uptake rate increased in the order of SCN^- < SO₄^2-≈Cl^- < HPO₄^2-. Dynamic binding capacity increased in the order of SCN^- < SO₄^2- < HPO₄^2- < Cl^-, the same of the order of adsorption capacity. The dynamic binding capacity kept the highest values when Cl^- existed at the two ionic strengths. Protein elution was little affected by counterions. The results indicated that the adsorption performance of FF-PEI-R440 was affected by counterions and the Cl^- was the most favorable couterion for column operation.

Key words: ion exchange chromatography, polymer grafting, counterion, adsorption equilibrium, kinetics, dynamic binding capacity, linear gradient elution

中图分类号:

TQ033

翟秋红, 余林玲, 孙彦. 反离子对部分电荷中和的聚乙烯亚胺接枝介质的蛋白质色谱行为的影响[J]. 化工学报, 2017, 68(11): 4178-4185.

ZHAI Qiuhong, YU Linling, SUN Yan. Effect of counterions on protein adsorption to partial neutralization poly(ethylenimine)-grafted Sepharose FF[J]. CIESC Journal, 2017, 68(11): 4178-4185.

参考文献 31

[1]	RICCARDI E, WANG J C, LIAPIS A I. Modeling the construction of polymeric adsorbent media:effects of counter-ions on ligand immobilization and pore structure[J]. Journal of Chemical Physics, 2014, 140(8):3073-3083.
[2]	YU L L, ZHANG L, SUN Y. Protein behavior at surfaces:orientation, conformational transitions and transport[J]. Journal of Chromatography A, 2015, 1382:118-134.
[3]	LENHOFF A M. Protein adsorption and transport in polymer-functionalized ion-exchangers[J]. Journal of Chromatography A, 2011, 1218(49):8748-8759.
[4]	BASCONI J E, CARTA G, SHIRTS M R. Multiscale modeling of protein adsorption and transport in macroporous and polymer-grafted ion exchangers[J]. AIChE Journal, 2014, 60(11):3888-3901.
[5]	TAO Y Y, CHEN N Y, CARTA G, et al. Modeling multicomponent adsorption of monoclonal antibody charge variants in cation exchange columns[J]. AIChE Journal, 2012, 58(8):2503-2511.
[6]	余林玲, 孙彦. 接枝聚合物配基的蛋白质吸附层析[J]. 化工学报, 2016, 67(1):140-151. YU L L, SUN Y. Adsorptive protein chromatography with grafted polymeric ligands[J]. CIESC Journal, 2016, 67(1):140-151.
[7]	YU L L, TAO S P, DONG X Y, et al. Protein adsorption to poly(ethylenimine)-modified Sepharose FF (Ⅰ):A critical ionic capacity for drastically enhanced capacity and uptake kinetics[J]. Journal of Chromatography A, 2013, 1305(1):76-84.
[8]	HONG Y, LIU N, WEI W, et al. Protein adsorption to poly(ethylenimine)-modified Sepharose FF (Ⅲ):Comparison between different proteins[J]. Journal of Chromatography A, 2014, 1342(4):30-36.
[9]	YU L L, SUN Y. Protein adsorption to poly(ethylenimine)-modified Sepharose FF (Ⅱ):Effect of ionic strength[J]. Journal of Chromatography A, 2013, 1305(1):85-93.
[10]	TAO Y, PEREZ-ALMODOVAR E X, CARTA G, et al. Adsorption kinetics of deamidated antibody variants on macroporous and dextran-grafted cation exchangers (Ⅲ):Microscopic studies[J]. Journal of Chromatography A, 2011, 1218(44):8027-8035.
[11]	PEREZ-ALMODOVAR E X, WU Y G, CARTA G. Multicomponent adsorption of monoclonal antibodies on macroporous and polymer grafted cation exchangers[J]. Journal of Chromatography A, 2012, 1264(22):48-56.
[12]	ZHAO Y Y, DONG X Y, YU L L, et al. Protein adsorption to poly(ethylenimine)-modified Sepharose FF (Ⅵ):Partial charge neutralization drastically increases uptake rate[J]. Journal of Chromatography A, 2015, 1427:102-110.
[13]	STÅHLBERG J. Retention models for ions in chromatography 1[J]. Journal of Chromatography A, 1999, 855(1):3-55.
[14]	FRITZ J S. Factors affecting selectivity in ion chromatography[J]. Journal of Chromatography A, 2005, 1085(1):8-17.
[15]	TUGCU N, SONG M, BRENEMAN C M, et al. Prediction of the effect of mobile-phase salt type on protein retention and selectivity in anion exchange systems[J]. Analytical Chemistry, 2003, 75(14):3563-3572.
[16]	LIU N, YU L L, SUN Y. Protein adsorption to poly(ethylenimine)-modified Sepharose FF (Ⅴ):Complicated effects of counterions[J]. Journal of Chromatography A, 2015, 1404:44-50.
[17]	LIU N, WANG Z Y, LIU X G, et al. Characterization of novel mixed-mode protein adsorbents fabricated from benzoyl-modified polyethyleneimine-grafted Sepharose[J]. Journal of Chromatography A, 2014, 1372:157-165.
[18]	YONG J L, NOTIDES A C, TSAY Y G, et al. Coumestrol, NBD-norhexestrol, and dansyl-norhexestrol, fluorescent probes of estrogen-binding proteins[J]. Biochemistry, 1977, 16(13):2896-2901.
[19]	?MIECHOWSKI M. Theoretical calculation of pKas of phosphoric (Ⅴ) acid in the polarisable continuum and cluster-continuum models[J]. Journal of Molecular Structure, 2009, s924/925/926:170-174.
[20]	MARCUS Y. Thermodynamics of solvation of ions (Ⅴ):Gibbs free energy of hydration at 298. 15 K[J]. Journal of the Chemical Society Faraday Transactions, 1991, 87(18):2995-2999.
[21]	CARTA G, UBIERA A R, PABST T M. Protein mass transfer kinetics in ion exchange media:measurements and interpretations[J]. Chemical Engineering & Technology, 2005, 28(11):1252-1264.
[22]	LI M, LI Y Y, YU L L, et al. Characterization of poly (allylamine) as a polymeric ligand for ion-exchange protein chromatography[J]. Journal of Chromatography A, 2017, 1486:103-109.
[23]	ANGELO J M, CVETKOVIC A, GANTIER R, et al. Characterization of cross-linked cellulosic ion-exchange adsorbents (Ⅰ):Structural properties[J]. Journal of Chromatography A, 2013, 1319(18):46-56.
[24]	TIONG W H C, DAMODARAN G, NAIK H, et al. Enhancing amine terminals in an amine-deprived collagen matrix[J]. Langmuir, 2008, 24(20):11752-11761.
[25]	MADISON S A, CARNALI J O. pH Optimization of amidation via carbodiimides[J]. Industrial & Engineering Chemistry Research, 2013, 52(38):13547-13555.
[26]	CACACE M G, LANDAU E M, RAMSDEN J J. The Hofmeister series:salt and solvent effects on interfacial phenomena[J]. Quarterly Reviews of Biophysics, 1997, 30(3):241-277.
[27]	PEREZ-ALMODOVAR E X, GLATZ B, CARTA G. Counterion effects on protein adsorption equilibrium and kinetics in polymer-grafted cation exchangers[J]. Journal of Chromatography A, 2012, 1253(1253):83-93.
[28]	STONE M C, TAO Y, CARTA G. Protein adsorption and transport in agarose and dextran-grafted agarose media for ion exchange chromatography:effect of ionic strength and protein characteristics[J]. Journal of Chromatography A, 2009, 1216(20):4465-4474.
[29]	HU X, DO D D, YU Q. Effects of supporting and buffer electrolytes (NaCl, CH3COOH and NH4OH) on the diffusion of BSA in porous media[J]. Chemical Engineering Science, 1992, 47(1):151-164.
[30]	MÜLLER E. Properties and characterization of high capacity resins for biochromatography[J]. Chemical Engineering & Technology, 2005, 28(11):1295-1305.
[31]	FRANKE A, FORRER N, BUTTÉ A, et al. Role of the ligand density in cation exchange materials for the purification of proteins[J]. Journal of Chromatography A, 2010, 1217(15):2216-2225.

反离子对部分电荷中和的聚乙烯亚胺接枝介质的蛋白质色谱行为的影响

Effect of counterions on protein adsorption to partial neutralization poly(ethylenimine)-grafted Sepharose FF

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献 31

相关文章 15

编辑推荐

Metrics

本文评价

[1]	金正浩, 封立杰, 李舒宏. 氨水溶液交叉型再吸收式热泵的能量及分析[J]. 化工学报, 2023, 74(S1): 53-63.
[2]	程成, 段钟弟, 孙浩然, 胡海涛, 薛鸿祥. 表面微结构对析晶沉积特性影响的格子Boltzmann模拟[J]. 化工学报, 2023, 74(S1): 74-86.
[3]	肖明堃, 杨光, 黄永华, 吴静怡. 浸没孔液氧气泡动力学数值研究[J]. 化工学报, 2023, 74(S1): 87-95.
[4]	毕丽森, 刘斌, 胡恒祥, 曾涛, 李卓睿, 宋健飞, 吴翰铭. 粗糙界面上纳米液滴蒸发模式的分子动力学研究[J]. 化工学报, 2023, 74(S1): 172-178.
[5]	于宏鑫, 邵双全. 水结晶过程的分子动力学模拟分析[J]. 化工学报, 2023, 74(S1): 250-258.
[6]	范孝雄, 郝丽芳, 范垂钢, 李松庚. LaMnO₃/生物炭催化剂低温NH₃-SCR催化脱硝性能研究[J]. 化工学报, 2023, 74(9): 3821-3830.
[7]	郑佳丽, 李志会, 赵新强, 王延吉. 离子液体催化合成2-氰基呋喃反应动力学研究[J]. 化工学报, 2023, 74(9): 3708-3715.
[8]	汪林正, 陆俞冰, 张睿智, 罗永浩. 基于分子动力学模拟的VOCs热氧化特性分析[J]. 化工学报, 2023, 74(8): 3242-3255.
[9]	杨越, 张丹, 郑巨淦, 涂茂萍, 杨庆忠. NaCl水溶液喷射闪蒸-掺混蒸发的实验研究[J]. 化工学报, 2023, 74(8): 3279-3291.
[10]	曾如宾, 沈中杰, 梁钦锋, 许建良, 代正华, 刘海峰. 基于分子动力学模拟的Fe₂O₃纳米颗粒烧结机制研究[J]. 化工学报, 2023, 74(8): 3353-3365.
[11]	李锦潼, 邱顺, 孙文寿. 煤浆法烟气脱硫中草酸和紫外线强化煤砷浸出过程[J]. 化工学报, 2023, 74(8): 3522-3532.
[12]	张蒙蒙, 颜冬, 沈永峰, 李文翠. 电解液类型对双离子电池阴阳离子储存行为的影响[J]. 化工学报, 2023, 74(7): 3116-3126.
[13]	何宣志, 何永清, 闻桂叶, 焦凤. 磁液液滴颈部自相似破裂行为[J]. 化工学报, 2023, 74(7): 2889-2897.
[14]	李艳辉, 丁邵明, 白周央, 张一楠, 于智红, 邢利梅, 高鹏飞, 王永贞. 非常规服役超临界锅炉的微纳尺度腐蚀动力学模型建立及应用[J]. 化工学报, 2023, 74(6): 2436-2446.
[15]	王光宇, 张锴, 张凯华, 张东柯. 微波加热干燥煤泥热质传递及其能耗特性分析[J]. 化工学报, 2023, 74(6): 2382-2390.