Preparation and characterization of prototypes for multi-modal separation media aimed for capture of negatively charged biomolecules at high salt conditions

Johansson, Bo‐Lennart; Belew, Makonnen; Eriksson, Stefan; Glad, Gunnar; Lind, Ola; Maloisel, Jean-Luc; Norrman, Nils

doi:10.1016/s0021-9673(03)01140-3

Cited by 107 publications

(86 citation statements)

References 19 publications

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“…This allowed the direct determination of how charge density affects overall transport, retention and capacity while eliminating adsorbent structural differences, and together with previously reported results, it allows definition and prediction of the onset of the change in mass transport. Past charge density studies have predominantly been outside the field of ion-exchange adsorbents [28,29], but work on ion-exchange adsorbents found that the overall adsorbent capacity did not increase for charge densities above a specific threshold, while retention did increase with increasing ligand density [30]. However, transport effects were not evaluated in that work.…”

Section: Introductionmentioning

confidence: 97%

Chromatography of proteins on charge-variant ion exchangers and implications for optimizing protein uptake rates

Langford

Yan

et al. 2007

Journal of Chromatography A

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Intraparticle transport of proteins usually represents the principal resistance controlling their uptake in preparative separations. In ion-exchange chromatography two limiting models are commonly used to describe such uptake: pore diffusion, in which only free protein in the pore lumen contributes to transport, and homogeneous diffusion, in which the transport flux is determined by the gradient in the total protein concentration, free or adsorbed. Several studies have noted a transition from pore to homogeneous diffusion with increasing ionic strength in some systems, and here we investigate the mechanistic basis for this transition. The studies were performed on a set of custom-synthesized methacrylate-based strong cation exchangers differing in ligand density into which uptake of two proteins was examined using confocal microscopy and frontal loading experiments. We find that the transition in uptake mechanism occurs in all cases studied, and generally coincides with an optimum in the dynamic binding capacity at moderately high flow rates. The transition appears to occur when protein-surface attraction is weakened sufficiently, and this is correlated with the isocratic retention factor k' for the system of interest: the transition occurs in the vicinity of k' ~ 3000. This result, which may indicate that adsorption is sufficiently weak to allow the protein to diffuse along or near the surface, provides a predictive basis for optimizing preparative separations using only isocratic retention data.

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Section: Introductionmentioning

confidence: 97%

Chromatography of proteins on charge-variant ion exchangers and implications for optimizing protein uptake rates

Langford

Yan

et al. 2007

Journal of Chromatography A

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“…Streamline Direct CST I is a new type of cation exchanger with multimodal functional groups, which not only takes advantage of electrostatic interaction, but also takes advantage of hydrogen bond interaction and hydrophobic interaction to tightly bind proteins. In other words, the new type of multi-modal ligand on adsorbent is able to interact with proteins through various intermolecular forces to get a high binding capacity in high ionic strength and salt concentration feedstocks (Johansson et al, 2003a(Johansson et al, , 2003b.…”

Section: Introductionmentioning

confidence: 99%

Expanded bed adsorption/desorption of proteins with Streamline Direct CST I adsorbent

Xiu²,

Mata³

et al. 2006

Biotech & Bioengineering

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Streamline Direct CST I is a new type of ion exchanger with multi-modal functional groups, specially designed for an expanded bed adsorption (EBA) process, which can capture directly the proteins from the high ionic strength feedstocks with a high binding capacity. In this study, an experimental study is carried out for two-component proteins (BSA and myoglobin) competitive adsorption and desorption in an expanded bed packed with Streamline Direct CST I. Based on the measurements of the single- and two-component bovine serum albumin (BSA)/myoglobin adsorption isotherm on Streamline Direct CST I, the binding and elution conditions for the whole EBA process are selected; and then frontal analysis for a longer timescale and column displacement experiments in a fixed bed (XK16/20 column) are carried out to evaluate the two-component proteins (BSA and myoglobin) competitive adsorption and displacement on Streamline Direct CST I. Finally, the feasibility of capturing both BSA and myoglobin by an expanded bed packed with Streamline Direct CST I is addressed in a Streamline 50 column packed with 300 mL Streamline Direct CST I.

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“…Recent advances in the design of multimodal (MM) chromatographic systems have produced previously undescribed classes of chromatographic materials that can provide alternative and improved selectivities as compared to traditional single mode chromatographic materials (3)(4)(5)(6)(7)(8)(9). Johansson et al have developed a library of MM ligands that can be employed for the capture of charged proteins under high salt conditions (4,5).…”

mentioning

confidence: 99%

“…Johansson et al have developed a library of MM ligands that can be employed for the capture of charged proteins under high salt conditions (4,5). Liu et al have developed a silica-based MM resin capable of weak anion-exchange and reversed-phase interactions for the simultaneous separation of acidic, basic, and neutral pharmaceutical compounds (9).…”

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confidence: 99%

Evaluation of protein adsorption and preferred binding regions in multimodal chromatography using NMR

Chung

Freed

Holstein

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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NMR titration experiments with labeled human ubiquitin were employed in concert with chromatographic data obtained with a library of ubiquitin mutants to study the nature of protein adsorption in multimodal (MM) chromatography. The elution order of the mutants on the MM resin was significantly different from that obtained by ion-exchange chromatography. Further, the chromatographic results with the protein library indicated that mutations in a defined region induced greater changes in protein affinity to the solid support. Chemical shift mapping and determination of dissociation constants from NMR titration experiments with the MM ligand and isotopically enriched ubiquitin were used to determine and rank the relative binding affinities of interaction sites on the protein surface. The results with NMR confirmed that the protein possessed a distinct preferred binding region for the MM ligand in agreement with the chromatographic results. Finally, coarsegrained ligand docking simulations were employed to study the modes of interaction between the MM ligand and ubiquitin. The use of NMR titration experiments in concert with chromatographic data obtained with protein libraries represents a previously undescribed approach for elucidating the structural basis of protein binding affinity in MM chromatographic systems.ligand binding site | mixed mode chromatography | protein-ligand interactions | binding site mapping | pseudoaffinity T he development of efficient bioseparation processes for the production of high-purity biopharmaceuticals is one of the most pressing challenges facing the pharmaceutical and biotechnology industries today. In addition, high-resolution separations for complex bioanalytical applications are becoming increasingly important. Although it is generally accepted that nonspecific interactions can often complicate single mode chromatographic separations (e.g., ion-exchange, reversed-phase), these additional interactions can also result in unexpected selectivities (1, 2).Recent advances in the design of multimodal (MM) chromatographic systems have produced previously undescribed classes of chromatographic materials that can provide alternative and improved selectivities as compared to traditional single mode chromatographic materials (3-9). Johansson et al. have developed a library of MM ligands that can be employed for the capture of charged proteins under high salt conditions (4, 5). Liu et al. have developed a silica-based MM resin capable of weak anion-exchange and reversed-phase interactions for the simultaneous separation of acidic, basic, and neutral pharmaceutical compounds (9). Small ligand pseudoaffinity chromatographic materials such as those used for hydrophobic charge induction chromatography have resulted in previously undescribed classes of MM ligands that offer unique selectivities due more to multiple low affinity MM interactions than to specific binding to certain classes of proteins (10). In addition, several libraries of MM ligands have been recently developed and employed on chro...

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Preparation and characterization of prototypes for multi-modal separation media aimed for capture of negatively charged biomolecules at high salt conditions

Cited by 107 publications

References 19 publications

Chromatography of proteins on charge-variant ion exchangers and implications for optimizing protein uptake rates

Chromatography of proteins on charge-variant ion exchangers and implications for optimizing protein uptake rates

Expanded bed adsorption/desorption of proteins with Streamline Direct CST I adsorbent

Evaluation of protein adsorption and preferred binding regions in multimodal chromatography using NMR

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