Protein adsorption onto diethylaminoethyl dextran modified anion exchanger: Effect of ionic strength and column behavior

doi:10.1016/j.cjche.2017.07.013

Chinese Journal of Chemical Engineering ›› 2018, Vol. 26 ›› Issue (2): 259-267.DOI: 10.1016/j.cjche.2017.07.013

• Separation Science and Engineering • 上一篇下一篇

Protein adsorption onto diethylaminoethyl dextran modified anion exchanger: Effect of ionic strength and column behavior

Shu Bai, Lingli Gong, Detao Han, Yutong Li, Linling Yu, Yan Sun

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

收稿日期:2017-03-04 修回日期:2017-07-21 出版日期:2018-02-28 发布日期:2018-03-16
通讯作者: Linling Yu
基金资助:
Supported by the National Natural Science Foundation of China (21406160, 21621004).

Protein adsorption onto diethylaminoethyl dextran modified anion exchanger: Effect of ionic strength and column behavior

Shu Bai, Lingli Gong, Detao Han, Yutong Li, Linling Yu, Yan Sun

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:2017-03-04 Revised:2017-07-21 Online:2018-02-28 Published:2018-03-16
Contact: Linling Yu
Supported by:
Supported by the National Natural Science Foundation of China (21406160, 21621004).

摘要/Abstract

摘要： Our previous studies on bovine serum albumin (BSA) adsorption to diethylaminoethyl dextran (DEAE dextran, DexD, grafting-ligand) and DEAE (D, surface-ligand) modified Sepharose FF resins found that all the grafted resins (FF-DexD and FF-D-DexD) exhibited extremely fast uptake rate (effective diffusivity, D_e, D_e/D₀N 1.4), which was six times greater than the ungrafted resins (D_e/D₀b 0.3). In this work, the influence of ionic strength (IS) on 6 typical DEAE dextran-grafted resins was investigated. Bath adsorption equilibria and kinetics, breakthrough, and linear gradient elution experiments were conducted. Commercial DEAE Sepharose FF was used for comparison. It is found that protein adsorption capacities on DEAE dextran-FF resins and the commercial resin decreased with increasing IS, but DEAE dextran-FF resins exhibited much higher capacity sensitivity to salt concentration. Besides, steeper decrease of adsorption capacities could be obtained at higher graftingligand or surface-ligand density. It is worth noting that the facilitating role of surface-ligand to the "chain delivery" effect was weakened after adding salt, leading to the less improvement in uptake rate by increasing surface-ligand density at higher IS. Although the uptake rates of the DEAE dextran-FF resins increased first and then decreased with increasing IS, they kept the extremely high level of D_e values (D_e/D₀N 1.1) at the their working/binding IS range. Moreover, the DEAE dextran-FF resin displayed much higher adsorption capacities and D_e values than commercial ungrafted resin in their working condition. Furthermore, the column results of DEAE dextran-FF resins presented higher dynamic binding capacities than and similar elution ISs with DEAE Sepharose FF to achieve similar (or even higher) recoveries suggest the excellent chromatographic column performance of the DEAE dextran-FF resins. Finally, both high recovery and purity of BSA and γ-globulin could be easily achieved using the typical DEAE dextran-FF column, FF-D60-DexD160, to separate their binary mixtures, by step gradient elution. The research has provided new insights into the practical application of the series of DEAE-dextran grafted resins in protein chromatography and proved their superiority.

关键词: Ion exchange chromatography, DEAE dextran, Protein adsorption, Uptake kinetics, Ionic strength

Abstract: Our previous studies on bovine serum albumin (BSA) adsorption to diethylaminoethyl dextran (DEAE dextran, DexD, grafting-ligand) and DEAE (D, surface-ligand) modified Sepharose FF resins found that all the grafted resins (FF-DexD and FF-D-DexD) exhibited extremely fast uptake rate (effective diffusivity, D_e, D_e/D₀N 1.4), which was six times greater than the ungrafted resins (D_e/D₀b 0.3). In this work, the influence of ionic strength (IS) on 6 typical DEAE dextran-grafted resins was investigated. Bath adsorption equilibria and kinetics, breakthrough, and linear gradient elution experiments were conducted. Commercial DEAE Sepharose FF was used for comparison. It is found that protein adsorption capacities on DEAE dextran-FF resins and the commercial resin decreased with increasing IS, but DEAE dextran-FF resins exhibited much higher capacity sensitivity to salt concentration. Besides, steeper decrease of adsorption capacities could be obtained at higher graftingligand or surface-ligand density. It is worth noting that the facilitating role of surface-ligand to the "chain delivery" effect was weakened after adding salt, leading to the less improvement in uptake rate by increasing surface-ligand density at higher IS. Although the uptake rates of the DEAE dextran-FF resins increased first and then decreased with increasing IS, they kept the extremely high level of D_e values (D_e/D₀N 1.1) at the their working/binding IS range. Moreover, the DEAE dextran-FF resin displayed much higher adsorption capacities and D_e values than commercial ungrafted resin in their working condition. Furthermore, the column results of DEAE dextran-FF resins presented higher dynamic binding capacities than and similar elution ISs with DEAE Sepharose FF to achieve similar (or even higher) recoveries suggest the excellent chromatographic column performance of the DEAE dextran-FF resins. Finally, both high recovery and purity of BSA and γ-globulin could be easily achieved using the typical DEAE dextran-FF column, FF-D60-DexD160, to separate their binary mixtures, by step gradient elution. The research has provided new insights into the practical application of the series of DEAE-dextran grafted resins in protein chromatography and proved their superiority.

Key words: Ion exchange chromatography, DEAE dextran, Protein adsorption, Uptake kinetics, Ionic strength

Shu Bai, Lingli Gong, Detao Han, Yutong Li, Linling Yu, Yan Sun. Protein adsorption onto diethylaminoethyl dextran modified anion exchanger: Effect of ionic strength and column behavior[J]. Chinese Journal of Chemical Engineering, 2018, 26(2): 259-267.

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Protein adsorption onto diethylaminoethyl dextran modified anion exchanger: Effect of ionic strength and column behavior

Protein adsorption onto diethylaminoethyl dextran modified anion exchanger: Effect of ionic strength and column behavior

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