色胺混合模式层析介质制备及配基密度影响研究
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摘要
混合模式层析是一种新型的生物分离技术,在生物大分子药物的分离纯化中展现了良好的应用价值。本研究以色胺作为配基,制备了一系列不同配基密度的新型混合模式介质(4FF-Try)。对介质进行了成分结构表征,并通过静态吸附与吸附动力学考察了pH与盐对不同配基密度介质吸附性能的影响。结果表明,4FF-Try对于牛血清白蛋白的吸附呈现典型的pH依赖性及良好的耐盐吸附能力,并且吸附能力随着配基密度的增加,呈现"U"型变化,即存在最佳配基密度范围(60～80μmol/g),最高吸附容量为74.5 mg/g。本研究结果证实了4FF-Try用于生物分离的潜力,可应用于介质制备过程与分离过程的优化。
Mixed mode chromatography is a new biological separation technology, which has shown good application on biomacromolecular purification. In this study, tryptamine was used as the ligand to prepare a series of mixed mode resin(4 FF-Try) with different ligand densities. The composition and structure of resin were characterized, and the effects of pH and salt on the adsorption performance were investigated by batch adsorption and adsorption kinetics. The results showed that the adsorption of resin toward BSA was pHdependent and salt-tolerant. In addition, with the increase of ligand density, the adsorption performance of4 FF-Try presented an U-shaped change. There was an optimal ligand density range(60～80 μmol/g) for 4 FFTry, and the highest adsorption capacity was 74.5 mg/g resin. The results of this study confirmed the potential of the new mixed mode resin 4 FF-Try for bioseparation, which could be applied to improve the resin preparation process and the separation process.
Mixed mode chromatography is a new biological separation technology, which has shown good application on biomacromolecular purification. In this study, tryptamine was used as the ligand to prepare a series of mixed mode resin(4 FF-Try) with different ligand densities. The composition and structure of resin were characterized, and the effects of pH and salt on the adsorption performance were investigated by batch adsorption and adsorption kinetics. The results showed that the adsorption of resin toward BSA was pHdependent and salt-tolerant. In addition, with the increase of ligand density, the adsorption performance of4 FF-Try presented an U-shaped change. There was an optimal ligand density range(60～80 μmol/g) for 4 FFTry, and the highest adsorption capacity was 74.5 mg/g resin. The results of this study confirmed the potential of the new mixed mode resin 4 FF-Try for bioseparation, which could be applied to improve the resin preparation process and the separation process.
引文
[1]Santarelli X. and Cabanne C., Mixed Mode Chromatography:A Novel Way Toward New Selectivity.[J]Curr. Protein Pept. Sci., 2019, 20.
[2]Zhang K. and Liu X.D., Mixed-mode chromatography in pharmaceutical and biopharmaceutical applications.[J] J. Pharmaceut. Biomed., 2016, 130.
[3]Halan V., Maity S., Bhambure R., and Rathore A.S., Multimodal Chromatography for Purification of Biotherapeutics-A Review.[J] Curr. Protein Pept. Sci., 2019, 20.
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[5]Tong H.F., Cavallotti C., Yao S.J., and Lin D.Q., Molecular insight into protein binding orientations and interaction modes on hydrophobic charge-induction resin.[J] J. Chromatogr. A, 2017, 1512.
[6]Lu H.L., Lin D.Q., Zhang Q.L., and Yao S.J., Evaluation on adsorption selectivity of immunoglobulin G with 2-mercapto-l-methyl-imidazole-based hydrophobic charge-induction resins.[J]Biochem. Eng. J., 2017, 119.
[7]Xia H.F., Lin D.Q., Wang L.P., Chen Z.J., and Yao S.J., Preparation and Evaluation of Cellulose Adsorbents for Hydrophobic Charge Induction Chromatography.[J] Ind. Eng. Chem. Res., 2008, 47.
[8]Shi W., Lin D.Q., Tong H.F., Yun J.X., and Yao S.J., 5-Aminobenzimidazole as new hydrophobic charge-induction ligand for expanded bed adsorption of bovine IgG.[J] J. Chromatogr. A, 2015, 1425.
[9]Cheng F., Li M., He W., Sun B., Qin J., and Qu J., Activation of resin with controllable ligand density via catalytic oxa-Michael addition and application in antibody purification.[J] J. Chromatogr. A, 2018, 1570.
[10]Bhambure R., Angelo J.M., Gillespie C.M., Phillips M., Graalfs H., and Lenhoff A.M., Ionic strength-dependent changes in tentacular ion exchangers with variable ligand density. II. Functional properties.[J] J. Chromatogr. A, 2017, 1506.
[11]Wang C.X., Lin D.Q., Liu T., and Yao S.J., Hydrophobic charge-induction chromatographic resin with 5-aminobenzimidazol ligand:Effects of ligand density on protein adsorption.[J] Sep. Sci. Technol., 2016, 51.
[12]Zhao W.W., Shi Q.H., and Sun Y., FYWHCLDE-based affinity chromatography of IgG:Effect of ligand density and purifications of human IgG and monoclonal antibody.[J] J. Chromatogr. A, 2014, 1355.
[13]Lu H.L., Lin D.Q., Zhu M.M., and Yao S.J., Protein adsorption on DEAE ion-exchange resins with different ligand densities and pore sizes.[J] J. Sep. Sci., 2012, 35.
[14]Lu H.L., Lin D.Q., Zhu M.M., and Yao S.J., Effects of ligand density and pore size on the adsorption of bovine IgG with DEAE ion-exchange resins.[J] J. Sep. Sci., 2012, 35.
[15]Rafferty J.L., Siepmann J.I., and Schure M.R., Influence of bonded-phase coverage in reversed-phase liquid chromatography via molecular simulation I. Effects on chain conformation and interfacial properties.[J] J.Chromatogr. A, 2008, 1204.
[16]Cheng F., Li M.-Y., Wang H.-Q., Lin D.-Q., and Qu J.-P., Antibody-Ligand Interactions for Hydrophobic Charge-Induction Chromatography:A Surface Plasmon Resonance Study.[J] Langmuir, 2015, 31.
[17]Wu Q.C., Lin D.Q., Shi W., Zhang Q.L., and Yao S.J., A mixed-mode resin with tryptamine ligand for human serum albumin separation.[J] J. Chromatogr. A, 2016, 1431.
[18]Shi W., Zhang S.-Q., Li K.-B., Jia W.-P., and Han D.-M., Integration of mixed-mode chromatography and molecular imprinting technology for double recognition and selective separation of proteins.[J] Sep.Purif. Technol., 2018, 202.
[19]Liu T., Lin D.Q., Lu H.L., and Yao S.J., Preparation and evaluation of dextran-grafted agarose resin for hydrophobic charge-induction chromatography.[J] J. Chromatogr. A, 2014, 1369.
[20]Yu L.L. and Sun Y., Protein adsorption to poly(ethylenimine)-modified Sepharose FF:II. Effect of ionic strength.[J] J. Chromatogr. A, 2013, 1305.
[21]Liu N., Wang Z., Liu X.M., Yu L., and Sun Y., Characterization of novel mixed-mode protein adsorbents fabricated from benzoyl-modified polyethyleneimine-grafted Sepharose.[J] J. Chromatogr. A, 2014, 1372.
[22]Yu L.L., Liu N., Hong Y., and Sun Y., Protein adsorption and chromatography on novel mixed-mode resins fabricated from butyl-modified poly(ethylenimine)-grafted Sepharose.[J] Chem. Eng. Sci., 2015, 135.
[2]Zhang K. and Liu X.D., Mixed-mode chromatography in pharmaceutical and biopharmaceutical applications.[J] J. Pharmaceut. Biomed., 2016, 130.
[3]Halan V., Maity S., Bhambure R., and Rathore A.S., Multimodal Chromatography for Purification of Biotherapeutics-A Review.[J] Curr. Protein Pept. Sci., 2019, 20.
[4]Wang L.J., Wei W.L., Xia Z.N., Jie X., and Xia Z.Z.L., Recent advances in materials for stationary phases of mixed-mode high-performance liquid chromatography.[J] Trac-Trend. Anal. Chem., 2016, 80.
[5]Tong H.F., Cavallotti C., Yao S.J., and Lin D.Q., Molecular insight into protein binding orientations and interaction modes on hydrophobic charge-induction resin.[J] J. Chromatogr. A, 2017, 1512.
[6]Lu H.L., Lin D.Q., Zhang Q.L., and Yao S.J., Evaluation on adsorption selectivity of immunoglobulin G with 2-mercapto-l-methyl-imidazole-based hydrophobic charge-induction resins.[J]Biochem. Eng. J., 2017, 119.
[7]Xia H.F., Lin D.Q., Wang L.P., Chen Z.J., and Yao S.J., Preparation and Evaluation of Cellulose Adsorbents for Hydrophobic Charge Induction Chromatography.[J] Ind. Eng. Chem. Res., 2008, 47.
[8]Shi W., Lin D.Q., Tong H.F., Yun J.X., and Yao S.J., 5-Aminobenzimidazole as new hydrophobic charge-induction ligand for expanded bed adsorption of bovine IgG.[J] J. Chromatogr. A, 2015, 1425.
[9]Cheng F., Li M., He W., Sun B., Qin J., and Qu J., Activation of resin with controllable ligand density via catalytic oxa-Michael addition and application in antibody purification.[J] J. Chromatogr. A, 2018, 1570.
[10]Bhambure R., Angelo J.M., Gillespie C.M., Phillips M., Graalfs H., and Lenhoff A.M., Ionic strength-dependent changes in tentacular ion exchangers with variable ligand density. II. Functional properties.[J] J. Chromatogr. A, 2017, 1506.
[11]Wang C.X., Lin D.Q., Liu T., and Yao S.J., Hydrophobic charge-induction chromatographic resin with 5-aminobenzimidazol ligand:Effects of ligand density on protein adsorption.[J] Sep. Sci. Technol., 2016, 51.
[12]Zhao W.W., Shi Q.H., and Sun Y., FYWHCLDE-based affinity chromatography of IgG:Effect of ligand density and purifications of human IgG and monoclonal antibody.[J] J. Chromatogr. A, 2014, 1355.
[13]Lu H.L., Lin D.Q., Zhu M.M., and Yao S.J., Protein adsorption on DEAE ion-exchange resins with different ligand densities and pore sizes.[J] J. Sep. Sci., 2012, 35.
[14]Lu H.L., Lin D.Q., Zhu M.M., and Yao S.J., Effects of ligand density and pore size on the adsorption of bovine IgG with DEAE ion-exchange resins.[J] J. Sep. Sci., 2012, 35.
[15]Rafferty J.L., Siepmann J.I., and Schure M.R., Influence of bonded-phase coverage in reversed-phase liquid chromatography via molecular simulation I. Effects on chain conformation and interfacial properties.[J] J.Chromatogr. A, 2008, 1204.
[16]Cheng F., Li M.-Y., Wang H.-Q., Lin D.-Q., and Qu J.-P., Antibody-Ligand Interactions for Hydrophobic Charge-Induction Chromatography:A Surface Plasmon Resonance Study.[J] Langmuir, 2015, 31.
[17]Wu Q.C., Lin D.Q., Shi W., Zhang Q.L., and Yao S.J., A mixed-mode resin with tryptamine ligand for human serum albumin separation.[J] J. Chromatogr. A, 2016, 1431.
[18]Shi W., Zhang S.-Q., Li K.-B., Jia W.-P., and Han D.-M., Integration of mixed-mode chromatography and molecular imprinting technology for double recognition and selective separation of proteins.[J] Sep.Purif. Technol., 2018, 202.
[19]Liu T., Lin D.Q., Lu H.L., and Yao S.J., Preparation and evaluation of dextran-grafted agarose resin for hydrophobic charge-induction chromatography.[J] J. Chromatogr. A, 2014, 1369.
[20]Yu L.L. and Sun Y., Protein adsorption to poly(ethylenimine)-modified Sepharose FF:II. Effect of ionic strength.[J] J. Chromatogr. A, 2013, 1305.
[21]Liu N., Wang Z., Liu X.M., Yu L., and Sun Y., Characterization of novel mixed-mode protein adsorbents fabricated from benzoyl-modified polyethyleneimine-grafted Sepharose.[J] J. Chromatogr. A, 2014, 1372.
[22]Yu L.L., Liu N., Hong Y., and Sun Y., Protein adsorption and chromatography on novel mixed-mode resins fabricated from butyl-modified poly(ethylenimine)-grafted Sepharose.[J] Chem. Eng. Sci., 2015, 135.