Protein adsorption and chromatography on novel mixed-mode resins fabricated from butyl-modified poly(ethylenimine)-grafted Sepharose

Yu, Linling; Liu, Na; Hong, Yan; Sun, Yan

doi:10.1016/j.ces.2015.04.016

Cited by 19 publications

(5 citation statements)

References 41 publications

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“…In order to compare the groups fitted by different equations, Q m values would be replaced by the calculated adsorption density ( Q c ) to describe adsorption ability. Q c was introduced by Sun and co-workers − and derived by the fitting formula at a chosen equilibrium liquid-phase concentration. It was calculated by and where C C is a chosen equilibrium protein concentration in the liquid phase (mg/mL), Q m is the saturated adsorption capacity (mg/g resin), and K d is the dissociation constant (mg/mL).…”

Section: Resultsmentioning

confidence: 99%

Improvement of Adsorption Selectivity of Ion-Exchange Chromatography through the Double-Recognition Mode

Shi

Zhang

et al. 2019

J. Chem. Eng. Data

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Ion-exchange chromatography (IEC) was combined with molecular imprinting technology (MIT) to prepare two kinds of doublerecognition resins (DEAE@BSA and DEAE@bIgG) using bovine serum albumin (BSA) and bovine immunoglobulin G (bIgG) as the imprinting template proteins. New resins consist of ion-exchange interaction and spatial recognition and were characterized by Fourier transform infrared, scanning electron microscopy−energy-dispersive X-ray spectrometry, thermogravimetric analysis, and Brunauer−Emmett−Teller techniques. The static and dynamic adsorption behaviors of proteins on resins were investigated. In addition, the competitive adsorption between BSA and bIgG was also studied. It was found that the adsorbed protein density (Q c ) of the target protein remained the same after imprinting, but the competitive protein dropped obviously. The selectivity factors (α) of DEAE@BSA and DEAE@ bIgG could reach 3.56 and 2.56, respectively. The results of adsorption kinetics and frontal adsorption experiments further showed that the dynamic adsorption capacity of target proteins was improved in batch and column modes. Taken together, this study demonstrated that the selectivity of IEC could be improved with the double-recognition mode.

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Section: Resultsmentioning

confidence: 99%

Improvement of Adsorption Selectivity of Ion-Exchange Chromatography through the Double-Recognition Mode

Shi

Zhang

et al. 2019

J. Chem. Eng. Data

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“…Therefore, the fitted Q m values would be not suitable to describe the adsorption ability. As suggested by Sun and co-workers, − the adsorbed protein density ( Q c ) at an equilibrium liquid-phase concentration (5 mg/mL) calculated by Langmuir equation was used as a substitute to describe the adsorption capacity. The results are compared in Figure .…”

Section: Resultsmentioning

confidence: 99%

Mixed-Mode Expanded-Bed Adsorption for Human Serum Albumin Separation

Zhang

Dong

et al. 2018

Ind. Eng. Chem. Res.

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Mixed-mode chromatography (MMC) is a promising technology for protein separation with high adsorption capacity, good selectivity, salt-tolerance, and facile elution. Expanded bed adsorption (EBA) can capture target proteins directly from bioparticle containing crude feedstock without solid–liquid separation. In this study, MMC and EBA were combined to develop a mixed-mode EBA technique for human serum albumin (HSA) separation. Two mixed-mode EBA resins (using quartz-densified agarose beads and tungsten carbide/agarose composite beads, respectively) with tryptamine as the ligand were prepared (TA-S and TA-T). Static and dynamic adsorption behaviors of HSA were determined under varying conditions and typical adsorption properties of pH dependence and salt-tolerance were found. The performance of the resins in expanded bed was studied, and the dynamic binding capacities at 10% breakthrough (Q 10%) of TA-S and TA-T at bed expansion of 1.8 were 27.54 and 18.04 mg/mL settled resin, respectively. Moreover, these two resins were used to separate recombinant HSA (rHSA) from Pichia pastoris culture broth. TA-S showed better separation performance and the purity of rHSA monomer reached to 83.3% with the recovery of 91.6% through a two-step elution process. Total content of rHSA monomer and degraded fragment was 99.8%. The results demonstrate that the combination of MMC and EBA would be a potential platform for protein capture with fewer pretreatments and high process efficiency.

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“…It was found that some adsorption isotherms did not reach the plateaus, especially for high salt concentration, so the fitted Q m values would be not suitable to describe adsorption ability. As suggested by Sun and co-workers − the adsorbed protein density ( Q c ) at an equilibrium liquid-phase concentration calculated by the Langmuir equation was introduced. The Q c values at the equilibrium liquid-phase concentration of 5 mg/mL were calculated and are compared in Figure b.…”

Section: Results and Discussionmentioning

confidence: 99%

“…The adsorption data were fitted with the Langmuir equation, and the saturated adsorption capacity ( Q m ) and the dissociation constant ( K d ) were obtained. The adsorbed protein density ( Q c ) was calculated with the correlated Langmuir equation at the equilibrium protein concentration of 5 mg/mL, as suggested by Sun and co-workers. − …”

Section: Experimental Sectionmentioning

confidence: 99%

Integration of Expanded Bed Adsorption and Hydrophobic Charge-Induction Chromatography for Monoclonal Antibody Separation

Shi

Dong

Yao

et al. 2017

Ind. Eng. Chem. Res.

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The integration of expanded bed adsorption (EBA) with hydrophobic charge-induction chromatography (HCIC) affords a promising new technology to capture antibody from the complex feedstock. New EBA resin T-ABI was prepared with hydrophobic charge-induction ligand 5-aminobenzimidazole (ABI) and used to separate monoclonal antibody (mAb) from CHO cell culture broth. The static and dynamic adsorption behaviors of hIgG and mAb were investigated, and the typical properties of pH dependence and salt tolerance were found. High dynamic binding capacities at 10% breakthrough (18.1–21.4 mg/mL resin) were obtained even for high operation velocities (711–1203 cm/h) in an expanded bed. By optimization of loading pH, elution pH, and expansion factor, hIgG could be separated from the protein mixture (2 mg/mL hIgG and 10 mg/mL bovine serum albumin) with high efficiency. Finally, mAb was separated directly from the CHO cell culture broth with T-ABI EBA under optimized conditions (loading at pH 7.0, elution at pH 4.0 or 4.5, expansion factor of 2.0), and the purity reached 93.7–97.7% with the recovery of 72.6–79.4%. The results indicated that T-ABI resin was suitable to capture hIgG from the complicated feedstock. New separation process with T-ABI EBA showed a promising potential for antibody purification with high productivity and process efficiency, which combined the advantages of HCIC and EBA.

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Protein adsorption and chromatography on novel mixed-mode resins fabricated from butyl-modified poly(ethylenimine)-grafted Sepharose

Cited by 19 publications

References 41 publications

Improvement of Adsorption Selectivity of Ion-Exchange Chromatography through the Double-Recognition Mode

Improvement of Adsorption Selectivity of Ion-Exchange Chromatography through the Double-Recognition Mode

Mixed-Mode Expanded-Bed Adsorption for Human Serum Albumin Separation

Integration of Expanded Bed Adsorption and Hydrophobic Charge-Induction Chromatography for Monoclonal Antibody Separation

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