Our previous work proved that high adsorption capacity and uptake rate of lysozyme were achieved on alginate(Alg)-grafted re sin with an ionic capacity(IC) of 240 mmol·L^(-1)(Alg-FF-240).Moreover,the salt-toleran...Our previous work proved that high adsorption capacity and uptake rate of lysozyme were achieved on alginate(Alg)-grafted re sin with an ionic capacity(IC) of 240 mmol·L^(-1)(Alg-FF-240).Moreover,the salt-tolerant feature of Alg-FF-230 was improved by using sequential alginate grafting and sulfonation strategy.Inspired by the enhanced adsorption performance of lysozyme,we have herein proposed to investigate the static and dynamic adsorption behaviors of γ-globulin on a series of Alg-grafted resins with different grafting densities and sulfonation degrees.The adsorption ca pacity of γ-globulin decreased with increa sing alginate-grafting density(IC) from 160 to 230 mmol·L^(-1) at 0 mmol·L^(-1) NaCl because of the steric hindrance caused by the alginate-grafting layer.Effects of ionic strength(IS) indicated that the adsorption capacities of the resins with the IC value of 230-370 mmol·L^(-1) were much higher than CM Sepharose FF at 50-100 mmol·L^(-1) NaCl,and the uptake rate of Alg-FF-230 was about twice as much as that of CM Sepharose FF.This work demonstrated the important effects of alginate-grafting layer and IS in γ-globulin adsorption behavior,which would be helpful in the design of Alggrafted resins and the selection of proper IS condition for protein purification.展开更多
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...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 Na Cl. 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 over150 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.展开更多
基金supported by the National Natural Science Foundation of China (Nos. 21878222 and 21621004)。
文摘Our previous work proved that high adsorption capacity and uptake rate of lysozyme were achieved on alginate(Alg)-grafted re sin with an ionic capacity(IC) of 240 mmol·L^(-1)(Alg-FF-240).Moreover,the salt-tolerant feature of Alg-FF-230 was improved by using sequential alginate grafting and sulfonation strategy.Inspired by the enhanced adsorption performance of lysozyme,we have herein proposed to investigate the static and dynamic adsorption behaviors of γ-globulin on a series of Alg-grafted resins with different grafting densities and sulfonation degrees.The adsorption ca pacity of γ-globulin decreased with increa sing alginate-grafting density(IC) from 160 to 230 mmol·L^(-1) at 0 mmol·L^(-1) NaCl because of the steric hindrance caused by the alginate-grafting layer.Effects of ionic strength(IS) indicated that the adsorption capacities of the resins with the IC value of 230-370 mmol·L^(-1) were much higher than CM Sepharose FF at 50-100 mmol·L^(-1) NaCl,and the uptake rate of Alg-FF-230 was about twice as much as that of CM Sepharose FF.This work demonstrated the important effects of alginate-grafting layer and IS in γ-globulin adsorption behavior,which would be helpful in the design of Alggrafted resins and the selection of proper IS condition for protein purification.
基金supported by the National Natural Science Foundation of China (Nos. 21878222 and 21621004).
文摘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 Na Cl. 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 over150 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.
