Lysozyme adsorption to cation exchanger derivatized by sequential modification of poly(ethylenimine)-Sepharose with succinic anhydride and ethanolamine: Effect of pH and ionic strength

doi:10.1016/j.cjche.2019.07.003

Abstract

Abstract: In our previous work, a series of polyethylenimine (PEI)-derived cation exchangers were synthesized using PEIgrafted resin FF-PEI-L740 (ionic capacity, 740 mmol·L^-1) as the basic resin to study lysozyme adsorption and chromatographic behavior. It was found that the resin with an ionic capacity of 630 mmol·L^-1 (FF-PEI-CR630) possessed high adsorption performance towards lysozyme at 0-100 mmol·L^-1 NaCl. Therefore, in this work, FF-PEI-CR630 was selected to study the influences of pH and ionic strength (IS) on protein adsorption and chromatographic behavior towards lysozyme. The increase of lysozyme adsorption capacity in the pH range of 6 to 10 was observed. However, the uptake rate decreased in the pH range of 6 to 8 and then remained essentially unchanged from pH 8 to pH 10. Increasing IS led to decreased protein adsorption capacity and increased uptake rate in different pH ranges. Besides, FF-PEI-CR630 maintained dynamic binding capacity as high as over 150 mg·ml^-1 at pH 8-10 without NaCl. The research has thus provided insight into the selection of proper pH and IS conditions for protein purification by using FF-PEI-CR630.

Key words: Ion exchange, pH value, Adsorption, Kinetics, Chain delivery, Dynamic binding capacity

摘要： In our previous work, a series of polyethylenimine (PEI)-derived cation exchangers were synthesized using PEIgrafted resin FF-PEI-L740 (ionic capacity, 740 mmol·L^-1) as the basic resin to study lysozyme adsorption and chromatographic behavior. It was found that the resin with an ionic capacity of 630 mmol·L^-1 (FF-PEI-CR630) possessed high adsorption performance towards lysozyme at 0-100 mmol·L^-1 NaCl. Therefore, in this work, FF-PEI-CR630 was selected to study the influences of pH and ionic strength (IS) on protein adsorption and chromatographic behavior towards lysozyme. The increase of lysozyme adsorption capacity in the pH range of 6 to 10 was observed. However, the uptake rate decreased in the pH range of 6 to 8 and then remained essentially unchanged from pH 8 to pH 10. Increasing IS led to decreased protein adsorption capacity and increased uptake rate in different pH ranges. Besides, FF-PEI-CR630 maintained dynamic binding capacity as high as over 150 mg·ml^-1 at pH 8-10 without NaCl. The research has thus provided insight into the selection of proper pH and IS conditions for protein purification by using FF-PEI-CR630.

关键词: Ion exchange, pH value, Adsorption, Kinetics, Chain delivery, Dynamic binding capacity

Yangyang Zhao, Xianxiu Li, Linling Yu, Xiaoyan Dong, Yan Sun. Lysozyme adsorption to cation exchanger derivatized by sequential modification of poly(ethylenimine)-Sepharose with succinic anhydride and ethanolamine: Effect of pH and ionic strength[J]. Chinese Journal of Chemical Engineering, 2020, 28(2): 440-444.

References

[1] R. Bhambure, C.M. Gillespie, M. Phillips, H. Graalfs, A.M. Lenhoff, Ionic strengthdependent changes in tentacular ion exchangers with variable ligand density. I. Structural properties, J. Chromatogr. A 1463(2016) 90-101.
[2] Z. Yang, M. Gao, Z. Li, F. Zhang, S. Zhang, B. Yang, A poly(glycidylmethacrylatedivinylbenzene)-based anion exchanger for ion chromatography, J. Chromatogr. A 1596(2019) 79-83.
[3] Y. Zhao, X. Dong, L. Yu, Y. Liu, Y. Sun, Characterization of new polymer-grafted protein cation exchangers developed by partial neutralization of carboxyl groups derivatized by modification of poly(ethylenimine)-Sepharose with succinic anhydride, J. Chromatogr. A 1550(2018) 28-34.
[4] M. Li, Y. Wu, Y. Liu, Y. Sun, Protein adsorption to poly(allylamine)-modified Sepharose FF:Influences of polymer size and partial charge neutralization, Biochem. Eng. J. 142(2019) 153-161.
[5] X. Li, Y. Liu, Y. Sun, Alginate-grafted Sepharose FF:A novel polymeric ligand-based cation exchanger for high-capacity protein chromatography, Biochem. Eng. J. 126(2017) 50-57.
[6] A. Xue, L. Yu, Y. Sun, Implications from protein uptake kinetics onto dextran-grafted Sepharose FF coupled with ion exchange and affinity ligands, Chin. J. Chem. Eng. 25(2017) 906-910.
[7] L.L. Yu, S.P. Tao, X.Y. Dong, Y. Sun, Protein adsorption to poly(ethylenimine)-modified Sepharose FF:I. A critical ionic capacity for drastically enhanced capacity and uptake kinetics, J. Chromatogr. A 1305(2013) 76-84.
[8] Y. Hong, N. Liu, W. Wei, L.L. Yu, G. Ma, Y. Sun, Protein adsorption to poly (ethylenimine)-modified Sepharose FF:III. Comparison between different proteins, J. Chromatogr. A 1342(2014) 30-36.
[9] X. Li, Y. Liu, Y. Sun, Sequential alginate grafting and sulfonation significantly improve the performance of alginate-grafted Sepharose FF for protein chromatography, Biochem. Eng. J. 137(2018) 11-17.
[10] L. Ming, L. Yu, Y. Liu, Y. Sun, High uptake rate and extremely salt-tolerant behavior of protein adsorption to 900 kDa poly(allylamine)-modified Sepharose FF, Biochem. Eng. J. 134(2018) 94-100.
[11] Y. Zhao, X. Dong, L. Yu, Y. Sun, Protein adsorption to poly(ethylenimine)-modified Sepharose FF:VI. Partial charge neutralization drastically increases uptake rate, J. Chromatogr. A 1427(2016) 102-110.
[12] Y. Zhao, X. Dong, L. Yu, Y. Liu, Y. Sun, Implications from protein adsorption onto anion- and cation-exchangers derivatized by modification of poly(ethylenimine)-Sepharose FF with succinic anhydride, Biochem. Eng. J. 132(2018) 79-89.
[13] X. Li, Q. Wang, X. Dong, Y. Liu, Y. Sun, Grafting glycidyl methacrylate-iminodiacetic acid conjugate to Sepharose FF for fabrication of high-capacity protein cation exchangers, Biochem. Eng. J. 138(2018) 74-80.
[14] M.H.M. Olsson, C.R. Søndergaard, M. Rostkowski, J.H. Jensen, PROPKA3:Consistent treatment of internal and surface residues in empirical pKa predictions, J. Chem. Theory Comput. 7(2011) 525-537.
[15] K. Gekko, H. Noguchi, Potentiometric studies of hydrophobic effect on ion binding of ionic dextran derivatives, Biopolymers. 14(1975) 2555-2565.
[16] Y. Zhao, L. Yu, X. Dong, Y. Sun, Protein adsorption to poly(ethylenimine)-modified Sepharose FF:VII. Complicated effects of pH, J. Chromatogr. A 1580(2018) 72-79.
[17] Q. Wu, Q. Zhang, S. Xu, C. Ge, S. Yao, D. Lin, Preparation and evaluation of mixedmode resins with tryptophan analogues as functional ligands for human serum albumin separation, Chin. J. Chem. Eng. 25(2017) 898-905.
[18] M. Li, Y. Li, L. Yu, Y. Sun, Characterization of poly(allylamine) as a polymeric ligand for ion-exchange protein chromatography, J. Chromatogr. A 1486(2016) 103-109.
[19] M.C. Stone, Y. Tao, G. Carta, Protein adsorption and transport in agarose and dextran-grafted agarose media for ion exchange chromatography:Effect of ionic strength and protein characteristics, J. Chromatogr. A 1216(2009) 4465-4474.
[20] S.B. Dubin, G. Feher, G.B. Benedek, Study of the chemical denaturation of lysozyme by optical mixing spectroscopy, Biochemistry. 12(1973) 714-720.
[21] L.E. Weaver, G. Carta, Protein adsorption on cation exchangers:Comparison of macroporous and gel-composite media, Biotechnol. Prog. 12(2010) 342-355.
[22] Q.H. Shi, G.D. Jia, Y. Sun, Dextran-grafted cation exchanger based on superporous agarose gel:Adsorption isotherms, uptake kinetics and dynamic protein adsorption performance, J. Chromatogr. A 1217(2010) 5084-5091.
[23] N. Liu, L.L. Yu, Y. Sun, Protein adsorption to poly(ethylenimine)-modified Sepharose FF. IV. Dynamic adsorption and elution behaviors, J. Chromatogr. A 1362(2014) 218-224.
[24] X. Shan, W. Qin, Y. Dai, Dependence of extraction equilibrium of monocarboxylic acid from aqueous solutions on the relative basicity of extractant, Chem. Eng. Sci. 61(2006) 2574-2581.
[25] T.J. Su, J.R. Lu, R.K. Thomas, Z.F. Cui, J. Penfold, The effect of solution pH on the structure of lysosyme layers adsorbed at the silica-water interface studied by neutron reflection, Langmuir. 14(1998) 438-445.
[26] S.T. Moerz, P. Huber, pH-dependent selective protein adsorption into mesoporous silica, J. Phys. Chem. C 119(2015) 27072-27079.
[27] D. Gao, D.Q. Lin, S.J. Yao, Mechanistic analysis on the effects of salt concentration and pH on protein adsorption onto a mixed-mode adsorbent with cation ligand, J. Chromatogr. B 859(2007) 16-23.
[28] S. Wang, X. Li, Y. Sun, Poly(N,N-dimethylaminopropyl acrylamide)-grafted Sepharose FF:A new anion exchanger of very high capacity and uptake rate for protein chromatography, J. Chromatogr. A 1597(2019) 187-195.
[29] C. Harinarayan, J. Mueller, A. Ljungl, R. Fahrner, J.A. Van, R.R. Van, An exclusion mechanism in ion exchange chromatography, Biotechnol. Bioeng. 95(2010) 775-787.
[30] Y.M. Fang, D.Q. Lin, S.J. Yao, Adsorption of IgG and BSA on two chromatographic resins-Poly(ethylenimine)-4FF resin and tetrapeptide-poly(ethylenimine)-4FF resin, J. Chem. Eng. Data 63(2018) 4418-4424.
[31] H.Y. Wang, Y. Sun, S.L. Zhang, J. Luo, Q.H. Shi, H.Y. Wang, Y. Sun, S.L. Zhang, J. Luo, Q.H. Shi, Fabrication of high-capacity cation-exchangers for protein chromatography by atom transfer radical polymerization, Biochem. Eng. J. 113(2016) 19-29.
[32] L.L. Yu, Y. Sun, Protein adsorption to poly(ethylenimine)-modified Sepharose FF:II. Effect of ionic strength, J. Chromatogr. A 1305(2013) 85-93.
[33] S. Bai, L. Gong, D. Han, Y. Li, L. Yu, Y. Sun, Protein adsorption onto diethylaminoethyl dextran modified anion exchanger:Effect of ionic strength and column behavior, Chin. J. Chem. Eng. 26(2018) 259-267.
[34] B.D. Bowes, A.M. Lenhoff, Protein adsorption and transport in dextran-modified ionexchange media. II. Intraparticle uptake and column breakthrough, J. Chromatogr. A 1218(2011) 4698-4708.