Comparison of weak strong high-performance anion-exchange chromatography

Gooding, Karen M.; Schmuck, Mary Nell

doi:10.1016/s0021-9673(01)81642-3

Cited by 28 publications

(4 citation statements)

References 10 publications

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“…The contour plot of the log K values (Figure S4) demonstrated that the membrane operated in flowthrough mode across the majority of the binding conditions and no distinct strong and weak binding zone was observed. This may be due to complete ionization of the quaternary ammonium group (R‐CH 2 ‐N + ‐[CH 3 ] 3 ) on the Q membranes, a strong AEX ligand over broad pH ranges between pH 2 and pH 12 28 . The ionization of a weak ion exchange chromatography media varies with pH, whereas a strong ion exchanger remains fully ionized over broad pH range 29 .…”

Section: Resultsmentioning

confidence: 99%

“…This may be due to complete ionization of the quaternary ammonium group (R‐CH 2 ‐N + ‐[CH 3 ] 3 ) on the Q membranes, a strong AEX ligand over broad pH ranges between pH 2 and pH 12. 28 The ionization of a weak ion exchange chromatography media varies with pH, whereas a strong ion exchanger remains fully ionized over broad pH range. 29 In the case of the weak AEX such as Sartobind® STIC® PA, ionization of primary amine (R‐NH 2 ) ligand varies according to pH resulting in distinct binding zones.…”

Section: Resultsmentioning

confidence: 99%

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Design and optimization of membrane chromatography for monoclonal antibody charge variant separation

Nadar

Somasundaram

Charry

et al. 2022

Biotechnology Progress

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The manufacturing scale implementation of membrane chromatography to purify monoclonal antibodies has gradually increased with the shift in industry focus toward flexible manufacturing and disposable technologies. Membrane chromatography are used to remove process-related impurities such as host cell proteins (HCPs) and DNA, leachates, and endotoxins, with improved productivity and process flexibility.However, application of membrane chromatography to separate product-related variants such as charge variants has not gained major traction due to low-binding capacity. The work reported here demonstrates that a holistic process development strategy to optimize static binding (pH and salt concentration) and dynamic process (membrane loading, flowrate, and gradient length) parameters can alleviate the capacity limitations. The study employed high throughput screening tools and scale-down membranes for intermediate and polishing purification of the model monoclonal antibody. An optimized process consisting of anion exchange and cation exchange membrane chromatography reduced the acidic variants present in Protein A eluate from 89.5% to 19.2% with 71% recovery of the target protein. The membrane chromatography process also cleared HCP to below limit of detection with 6-to 30-fold higher membrane loading, compared to earlier reported values. The results confirm that membrane chromatography is effective in separating closely related product variants when supported by a well-defined process development strategy.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Design and optimization of membrane chromatography for monoclonal antibody charge variant separation

Nadar

Somasundaram

Charry

et al. 2022

Biotechnology Progress

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“…Experimental demonstration was on chromatography of oligosaccharides (Alm, 1952). Today, gradient elution techniques are widely used in analytical-and preparative-scale separation of proteins and other biochemicals (Lee et al, 1978;Yamamoto et al, 198313;Gooding and Schmuck, 1985). Many researchers have attempted to explain the behavior of gradient data through various models.…”

Section: Introductionmentioning

confidence: 99%

“…Goldberg (1985) presented evidence that intermediate forms exist in the protein folding process and that the unfolding process is often accompanied by the formation of aggregates. Gooding and Schmuck (1985) noted the appearance of multiple peaks for gradient elution of bovine serum albumin (BSA) under salt gradients in strong anionexchange HPLC. These peaks were attributed to aggregation of BSA.…”

Section: Introductionmentioning

confidence: 99%

Nonlinear Gradient Isotherm Parameter Estimation for Proteins with Consideration of Salt Competition and Multiple Forms

Whitley

Berninger

Rouhana

et al. 1991

Biotechnology Progress

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Salt gradients in ion-exchange chromatography are routinely used to speed separation of proteins and to concentrate products, but systematic optimization of these gradients requires protein equilibrium data as a function of salt concentration. An understanding of conformational changes, aggregation, and salt effects, which include both competition and affinity modulation, is important for equilibrium isotherm parameter estimation. In this study, gradient elution of bovine serum albumin (BSA) in anion exchange was well predicted by a salt-modulated nonlinear isotherm which considers salt competition. The isotherm was able to predict BSA gradient elution from batch equilibrium data. The same isotherm was also able to predict elution for various gradient slopes when fitted to an intermediate slope gradient experiment. If multiple forms due to aggregation or denaturation exist, isotherm parameters are readily averaged in batch experiments because of the long equilibration times. Similarly, gradient experiments yield averaged parameters because the salt gradient tends to merge the closely eluting forms. However, in isocractic elution, if the reaction rate is not rapid enough to give a merged peak, the estimated isotherm parameters are only fair predictors of gradient behavior and vice versa. Slower flow rates in isocratic elution can help reduce the discrepancy by allowing forms to merge through interconversion. As an alternative to determining averaged parameters, consideration of two binding forms, using VERSE-LC, an advanced rate model, gave good agreement with experimental data over the entire range of salt gradient durations.

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High‐Performance Liquid Chromatography of Peptides and Proteins

Boysen

Hearn

2011

Amino Acids, Peptides and Proteins in Organic Chemistry

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Comparison of weak strong high-performance anion-exchange chromatography

Cited by 28 publications

References 10 publications

Design and optimization of membrane chromatography for monoclonal antibody charge variant separation

Design and optimization of membrane chromatography for monoclonal antibody charge variant separation

Nonlinear Gradient Isotherm Parameter Estimation for Proteins with Consideration of Salt Competition and Multiple Forms

High‐Performance Liquid Chromatography of Peptides and Proteins

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