Mitigation of chromatography adsorbent lot performance variability through control of buffer solution design space

Aono, Hiromasa; Iliescu, Ionela; Cecchini, Doug; Wood, Susanne G.; McCue, Justin T.

doi:10.1016/j.chroma.2013.10.019

Cited by 7 publications

(2 citation statements)

References 8 publications

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“…While these prototypes varied in ligand density from 60-120 μmol/mL resin , previous work with Nuvia cPrime has shown that both protein selectivity and retention were relatively invariant with ligand density (R 2 = 0.93) over the range from 76-126 μmol/mL resin [22]. Thus, the variation in ligand density between the different prototypes was expected to have minimal effect on the retention behavior in these systems, an observation which has also been noted in several ion-exchange resin systems [28,29]. All of these ligands were immobilized on the same acrylamido gel matrix, which ensured an additional degree of comparability between the various resin materials.…”

Section: Assembling the Homologous Ligand Librarymentioning

confidence: 60%

Defining the property space for chromatographic ligands from a homologous series of mixed-mode ligands

Woo

Chen

Snyder

et al. 2015

Journal of Chromatography A

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A homologous ligand library based on the commercially-available Nuvia cPrime ligand was generated to systematically explore various features of a multimodal cation-exchange ligand and to identify structural variants that had significantly altered chromatographic selectivity. Substitution of the polar amide bond with more hydrophobic chemistries was found to enhance retention while remaining hydrophobically-selective for aromatic residues. In contrast, increasing the solvent exposure of the aromatic ring was observed to strengthen the ligand affinity for both types of hydrophobic residues. An optimal linker length between the charged and hydrophobic moieties was also observed to enhance retention, balancing the steric accessibility of the hydrophobic moiety with its ability to interact independently of the charged group. The weak pKa of the carboxylate charge group was found to have a notable impact on protein retention on Nuvia cPrime at lower pH, increasing hydrophobic interactions with the protein. Substituting the charged group with a sulfonic acid allowed this strong MM ligand to retain its electrostatic-dominant character in this lower pH range. pH gradient experiments were also carried out to further elucidate this pH dependent behavior. A single QSAR model was generated using this accumulated experimental data to predict protein retention across a range of multimodal and ion exchange systems. This model could correctly predict the retention of proteins on resins that were not included in the original model and could prove quite powerful as an in silico approach toward designing more effective and differentiated multimodal ligands.

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Section: Assembling the Homologous Ligand Librarymentioning

confidence: 60%

Defining the property space for chromatographic ligands from a homologous series of mixed-mode ligands

Woo

Chen

Snyder

et al. 2015

Journal of Chromatography A

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“…Special focus has been placed on the batch variations of IEX resins since it has been proven that it can play a significant role in the robustness of the downstream process [5,6]. Ligand density is known to be one of the main lot‐to‐lot variations, affecting protein adsorption and separation performance [5,7–9].…”

Section: Introductionmentioning

confidence: 99%

Mechanistic modeling of ligand density variations on anion exchange chromatography

Sanchez-Reyes

Graalfs

Hafner

et al. 2020

J of Separation Science

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Ion exchange chromatography is a powerful and ubiquitous unit operation in the purification of therapeutic proteins. However, the performance of an ion‐exchange process depends on a complex interrelationship between several parameters, such as protein properties, mobile phase conditions, and chromatographic resin characteristics. Consequently, batch variations of ion exchange resins play a significant role in the robustness of these downstream processing steps. Ligand density is known to be one of the main lot‐to‐lot variations, affecting protein adsorption and separation performance. The use of a model‐based approach can be an effective tool for comprehending the impact of parameter variations (e.g., ligand density) and their influence on the process. The objective of this work was to apply mechanistic modeling to gain a deeper understanding of the influence of ligand density variations in anion exchange chromatography. To achieve this, 13 prototype resins having the same support as the strong anion exchange resin Fractogel® EMD TMAE (M), but differing in ligand density, were analyzed. Linear salt gradient elution experiments were performed to observe the elution behavior of a monoclonal antibody and bovine serum albumin. A proposed isotherm model for ion exchange chromatography, describing the dependence of ligand density variations on protein retention, was successfully applied.

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Scaling Up Industrial Protein Chromatography

Antoniou

McCue

Natarajan

et al. 2017

Preparative Chromatography for Separation of Proteins

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Mitigation of chromatography adsorbent lot performance variability through control of buffer solution design space

Cited by 7 publications

References 8 publications

Defining the property space for chromatographic ligands from a homologous series of mixed-mode ligands

Defining the property space for chromatographic ligands from a homologous series of mixed-mode ligands

Mechanistic modeling of ligand density variations on anion exchange chromatography

Scaling Up Industrial Protein Chromatography

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