共聚物接枝蛋白离子交换色谱介质制备及性能

doi:10.11949/j.issn.0438-1157.20160349

化工学报 ›› 2016, Vol. 67 ›› Issue (9): 3738-3746.DOI: 10.11949/j.issn.0438-1157.20160349

共聚物接枝蛋白离子交换色谱介质制备及性能

张素玲, 杨葳, 余林玲, 白姝, 孙彦, 史清洪

天津大学化工学院生物化工系, 系统生物工程教育部重点实验室, 天津 300354

收稿日期:2016-03-28 修回日期:2016-05-09 出版日期:2016-09-05 发布日期:2016-09-05
通讯作者: 史清洪
基金资助:
国家自然科学基金项目（21476166，21236005）；天津市应用基础与前沿技术研究计划一般项目（15JCYBJC48500）。

Synthesis and characterization of copolymer-grafted cation-exchangers for protein adsorption

ZHANG Suling, YANG Wei, YU Linling, BAI Shu, SUN Yan, SHI Qinghong

Key Laboratory of System Bioengineering of Ministry of Education, Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300354, China

Received:2016-03-28 Revised:2016-05-09 Online:2016-09-05 Published:2016-09-05
Supported by:
supported by the National Natural Science Foundation of China (21476166, 21236005) and the Natural Science Foundation of Tianjin (15JCYBJC48500).

摘要/Abstract

摘要：

高容量蛋白质色谱介质是色谱过程高效化的材料基础和重要前提。采用原子转移自由基聚合（ATRP）技术，以甲基丙烯酸3-磺酸丙酯钾和甲基丙烯酸甲酯（MMA）为单体化合物合成了多种无规共聚物接枝离子交换色谱介质，并对其蛋白质吸附性能进行研究。单体总浓度一定情况下共聚物接枝色谱介质孔道半径（r_pore）随MMA浓度升高而增大，反映出接枝共聚物链渐趋塌陷的特征。蛋白质吸附结果表明，溶菌酶吸附容量取决于介质的离子交换容量；而抗体吸附容量则与rpore及相应的聚合物层厚度变化密切相关。随着聚合物层厚度的增大，聚合物层对抗体的空间排阻作用增强，抗体吸附容量下降。此外，引入MMA优化共聚物分子链可显著提高蛋白质吸附量，在SEP-gS30/M30介质中抗体和溶菌酶的饱和吸附量分别达到237 mg·g^-1和380 mg·g^-1。无规共聚物接枝离子交换色谱介质孔道内聚合物层厚度和蛋白质吸附也受无机盐浓度调控。

关键词: 共聚物接枝离子交换色谱介质, 原子转移自由基聚合, 蛋白质, 吸附, 高容量, 抗体

Abstract:

High-capacity chromatographic packings for protein adsorption are the material prerequisite for the development of efficient protein chromatography processes. In this work, random copolymer-grafted cation-exchangers for protein adsorption were synthesized by grafting 3-sulfopropyl methacrylate and methyl methacrylate (MMA) to Sepharose Fast Flow via atom transfer radical polymerization, and lysozyme and antibody adsorption on to these cation-exchangers was investigated. In case of a given total concentration of the monomers for the grafting reaction, the pore radii of the copolymer-grafted cation-exchangers increased with an increase of MMA concentration, indicating a typical characteristic of a decreasing depth of polymer layer and gradual collapse of the random copolymers. Adsorption equilibria showed that the adsorption capacity for lysozyme was dominated by ionic capacity of the cation-exchangers whilst for antibody it was related to pore radius and corresponding the change in depth of polymer layer. With an increase in depth of polymer layer, there was stronger steric excluded interaction between antibody and polymer layer, and thus decreasing the adsorption capacity for antibody. Moreover, the results demonstrated that the adsorption capacities for proteins on random copolymer-grafted cation-exchangers were improved by optimizing the conformation of random copolymer chains.The adsorption capacities for antibody and lysozyme reached 237 and 380 mg·g^-1 in the optimized random copolymer-grafted cation exchangers, respectively. The research revealed that both layer depth of copolymer and protein binding were also regulated by ionic strength in copolymer-grafted cation-exchangers.

Key words: copolymer-grafted cation-exchanger, atom transfer radical polymerization, protein, adsorption, high-capacity, antibody

中图分类号:

TQ936.2

张素玲, 杨葳, 余林玲, 白姝, 孙彦, 史清洪. 共聚物接枝蛋白离子交换色谱介质制备及性能[J]. 化工学报, 2016, 67(9): 3738-3746.

ZHANG Suling, YANG Wei, YU Linling, BAI Shu, SUN Yan, SHI Qinghong. Synthesis and characterization of copolymer-grafted cation-exchangers for protein adsorption[J]. CIESC Journal, 2016, 67(9): 3738-3746.

参考文献 23

[1]	AFEYAN N B, GORDON N F, MAZSAROFF I, et al. Flow-through particles for the high-performance liquid chromatographic separation of biomolecules:perfusion chromatography[J]. Journal of Chromatography, 1990, 519(1):1-29.
[2]	SHI Q H, ZHOU X, SUN Y. A novel superporous agarose medium for high-speed protein chromatography[J]. Biotechnology and Bioengineering, 2005, 92(5):643-651.
[3]	WANG D M, HAO G, SHI Q H, et al. Fabrication and characterization of superporous cellulose bead for high-speed protein chromatography[J]. Journal of Chromatography A, 2007, 1146(1):32-40.
[4]	SVEC F, FRECHET J M. New designs of macroporous polymers and supports:from separation to biocatalysis[J]. Science, 1996, 273(5272):205-211.
[5]	SVEC F. Porous polymer monoliths:amazingly wide variety of techniques enabling their preparation[J]. Journal of Chromatography A, 2010, 1217(6):902-924.
[6]	PARHI P, GOLAS A, BARNTHIP N, et al. Volumetric interpretation of protein adsorption:capacity scaling with adsorbate molecular weight and adsorbent surface energy[J]. Biomaterials, 2009, 30(36):6814-6824.
[7]	HORVATH J, BOSCHETTI E, GUERRIER L, et al. High-performance protein separations with novel strong ion exchangers[J]. Journal of Chromatography A, 1994, 679(1):11-22.
[8]	HAHN R, SCHULZ P M, SCHAUPP C, et al. Bovine whey fractionation based on cation-exchange chromatography[J]. Journal of Chromatography A, 1998, 795(2):277-287.
[9]	LENHOFF A M. Protein adsorption and transport in polymer-functionalized ion-exchangers[J]. Journal of Chromatography A, 2011, 1218(49):8748-8759.
[10]	LJUNGLOF A, LACKI K M, MUELLER J, et al. Ion exchange chromatography of antibody fragments[J]. Biotechnology and Bioengineering, 2007, 96(3):515-524.
[11]	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, 1305:76-84.
[12]	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, 1342:30-36.
[13]	YU L L, SUN Y. Protein adsorption to poly(ethylenimine)-modified Sepharose FF (Ⅱ):Effect of ionic strength[J]. Journal of Chromatography A, 2013, 1305:85-93.
[14]	王宏燕. 基于ATRP的聚合物接枝琼脂糖色谱介质合成及其性能表征[D]. 天津:天津大学, 2015. WANG H Y. Synthesis and characterization of agarose-based polymer-grafted chromatographic gels via atom transfer radical polymerization[D]. Tianjin:Tianjin University, 2015.
[15]	SHI Q H, JIA G D, SUN Y. Dextran-grafted cation exchanger based on superporous agarose gel:adsorption isotherms, uptake kinetics and dynamic protein adsorption performance[J]. Journal of Chromatography A, 2010, 1217(31):5084-5091.
[16]	YAO Y, LENHOFF A M. Pore size distributions of ion exchangers and relation to protein binding capacity[J]. Journal of Chromatography A, 2006, 1126(1/2):107-119.
[17]	SHI Q H, SHEN F F, SUN S. Studies of lysozyme binding to histamine as a ligand for hydrophobic charge induction chromatography[J]. Biotechnology Progress, 2010, 26(1):134-141.
[18]	ZHANG S P, SUN Y. Ionic strength dependence of protein adsorption to dye-ligand adsorbents[J]. AIChE Journal, 2002, 48(1):178-186.
[19]	SHI Q H, CHENG Z, SUN Y. 4-(1H-imidazol-1-yl)aniline:a new ligand of mixed-mode chromatography for antibody purification[J]. Journal of Chromatography A, 2009, 1216(33):6081-6087.
[20]	李海普, 李星, 张莎莎, 等. 高分子在固液界面吸附构象的研究方法及手段[J]. 化学通报, 2011, 74(5):417-423. LI H P, LI X, ZHANG S S, et al. Methods and techniques for studying the adsorption conformation of polymers at solid/liquid interface[J]. Chemistry Bulletin, 2011, 74(5):417-423.
[21]	DISMER F, PETZOLD M, HUBBUCH J. Effects of ionic strength and mobile phase pH on the binding orientation of lysozyme on different ion-exchange adsorbents[J]. Journal of Chromatography A, 2008, 1194(1):11-21.
[22]	DISMER F, HUBBUCH J. A novel approach to characterize the binding orientation of lysozyme on ion-exchange resins[J]. Journal of Chromatography A, 2007, 1149(2):312-320.
[23]	SHI Q H, JIA G D, XU L, et al. Effect of electric field on the partitioning behavior of solutes in entropic interaction chromatography[J]. Journal of Separation Science, 2013, 36(18):3075-3085.

共聚物接枝蛋白离子交换色谱介质制备及性能

Synthesis and characterization of copolymer-grafted cation-exchangers for protein adsorption

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