Displacement chromatography of proteins using a retained pH front in a hydrophobic charge induction chromatography column

Pinto, Nuno D.S.; Frey, Douglas D.

doi:10.1016/j.chroma.2015.01.087

Cited by 9 publications

(4 citation statements)

References 27 publications

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“…Хиболл-Нильсен и др. [67] Хроматография смещения -метод разделения веществ, который позволяет получить более высокие концентрации определенного компонента за счет вытеснения растворенным веществом, которое сорбируется интенсивнее, чем компоненты исходной смеси [77]. Принцип состоит в том, что образец загружается на верх колонки и перемещается с раствором, который является более сильным сорбентом, чем компоненты исходной смеси.…”

Section: нанофильтрация (нф)unclassified

Whey: State of the Art. Part II. Processes and Treatment Methods

Paladii¹,

Vrabie²,

Sprincean³

et al. 2021

EOM

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The current state of the art on studying of whey is considered. The processes and methods of whey processing (thermal, chemical, physicochemical, biotechnological, and electrophysical) are presented. Thermal and isoelectric precipitation of proteins, using reagents and coagulants, as well as the main membrane processing methods (reverse osmosis, diafiltration, microfiltration, ultrafiltration, and nanofiltration) are described. Possibi-lities of effective separation of whey proteins by a combination of membrane and other methods are noted. Chromatographic methods for fractionation of whey proteins (chromatography with imitation of a moving bed, high-gradient chromatography with a magnetic trap, selective adsorption, displacement chromatography, membrane adsorption), which provide a high degree of separation, are described. Highly porous chromatographic materials providing a high flow rate, biotechnological processing methods – biosynthesis of lactulose, enzymatic hydrolysis of lactose and whey proteins, aerobic and anaerobic fermentation are considered. Electrophysical methods of processing whey (electrodialysis, electroactivation), which include electrodialysis and electro-activation, as well as electrochemical activation as a phenomenon and technology, as a new promising processing method that allows to create a waste-free cycle for obtaining valuable components and useful derivatives from whey without the use of reagents are analyzed. It is emphasized that, depending on the regimes used, protein-mineral concentrates with a predetermined protein or mineral composition are obtained with the simultaneous isomerization of lactose into lactulose. It is stated that the efficiency of methods for processing whey is ensured by a significant increase in the efficiency of technological processes, a decrease in labor costs, a reduction in the processing time and materials, and an improvement in the quality and functional properties of the final products.

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Section: нанофильтрация (нф)unclassified

Whey: State of the Art. Part II. Processes and Treatment Methods

Paladii¹,

Vrabie²,

Sprincean³

et al. 2021

EOM

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“…The previous studies by Guo et al () and Pinto and Frey () extended the ion‐exchange equation‐based numerical model developed earlier by Frey et al () to mixed‐mode chromatography where the chromatographic ligands exhibit both electrostatic and hydrophobic interactions. These past efforts in particular highlighted the role of electrostatic interactions between buffering species and mixed‐mode chromatographic column packings in the formation of a retained pH gradient.…”

Section: Theorymentioning

confidence: 99%

Immunoglobulin G elution in protein A chromatography employing the method of chromatofocusing for reducing the co‐elution of impurities

Pinto

Uplekar

Moreira

et al. 2016

Biotech & Bioengineering

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Purification processes for monoclonal Immunoglobulin G (IgG) typically employ protein A chromatography as a capture step to remove most of the impurities. One major concern of the post-protein A chromatography processes is the co-elution of some of the host cell proteins (HCPs) with IgG in the capture step. In this work, a novel method for IgG elution in protein A chromatography that reduces the co-elution of HCPs is presented where a two-step pH gradient is self-formed inside a protein A chromatography column. The complexities involved in using an internally produced pH gradient in a protein A chromatography column employing adsorbed buffering species are discussed though equation-based modeling. Under the conditions employed, ELISA assays show a 60% reduction in the HCPs co-eluting with the IgG fraction when using the method as compared to conventional protein A elution without affecting the IgG yield. Evidence is also obtained which indicates that the amount of leached protein A present in free solution in the purified product is reduced by the new method. Biotechnol. Bioeng. 2017;114: 154-162. © 2016 Wiley Periodicals, Inc.

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“…In recent years, mixed-mode chromatography (MMC) was introduced as a novel technology for protein separation, which combines multiple binding modes, like ionic exchange, hydrophobic interaction, hydrogen bonding, etc. Good selectivity, high capacity, salt-tolerance and relatively low cost make MMC suitable for primary capture process. − Therefore, EBA and MMC, with their advantages, have been combined to develop a new protein capture technology, mixed-mode EBA, which has the potential to improve the process efficiency and avoid the frustrating pretreatments on the feedstock, such as clarification (solid–liquid separation), dilution (reducing the conductivity for ion exchange) and salt-adjustment (salt addition to enhance the hydrophobic interactions).…”

Section: Introductionmentioning

confidence: 99%

“…Hydrophobic charge-induction chromatography (HCIC), as one special MMC, was introduced for antibody purification, − which can bind antibody through hydrophobic interactions at neutral pH and elute effectively by electrostatic repulsion at acidic pH . The increasing clinical demand of monoclonal antibodies (mAbs) has promoted the revolution of antibody production techniques, and HCIC was developed as a cost-effective alternative to Protein A-based affinity chromatography for the capture of antibodies.…”

Section: Introductionmentioning

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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Displacement chromatography of proteins using a retained pH front in a hydrophobic charge induction chromatography column

Cited by 9 publications

References 27 publications

Whey: State of the Art. Part II. Processes and Treatment Methods

Whey: State of the Art. Part II. Processes and Treatment Methods

Immunoglobulin G elution in protein A chromatography employing the method of chromatofocusing for reducing the co‐elution of impurities

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

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