Semi-preparative high-performance reversed-phase displacement chromatography of insulins

Vigh, Gy.; Varga‐Puchony, Z.; Szepesi, G.; Gazdag, M.

doi:10.1016/s0021-9673(01)94612-6

Cited by 42 publications

(3 citation statements)

References 16 publications

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“…Displacement chromatography has emerged as a powerful technique for the purification of biomolecules that provides high product concentration, purity, and yield. – In this chromatographic separation process, the addition of a solute (a displacer) to the mobile phase enables the separation of components (e.g., proteins) that might otherwise have similar migration times. In particular, displacers with low molecular weight (<2000) offer distinct advantages for protein purification; – they can be readily separated from the purified protein, can be relatively cheap, and can display salt-dependent adsorption behavior facilitating column regeneration …”

Section: Introductionmentioning

confidence: 99%

An Affinity-Based Strategy for the Design of Selective Displacers for the Chromatographic Separation of Proteins

et al. 2008

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We describe an affinity-based strategy for designing selective protein displacers for the chromatographic purification of proteins. To design a displacer that is selective for a target protein, we attached a component with affinity for the target protein to a resin-binding component; we then tested the ability of such displacers to selectively retain the target protein on a resin relative to another protein having a similar retention time. In particular, we synthesized displacers based on biotin, which selectively retained avidin as compared to aprotinin on SP Sepharose high performance resin. In addition, we have extended this approach to develop an affinity-peptide-based displacer that discriminates between lysozyme and cytochrome c. Here, a selective displacer was designed from a lysozyme-binding peptide that had been identified and optimized previously using phage-display technology. Our results suggest a general strategy for designing highly selective affinity-based displacers by identifying molecules (e.g., peptides) that bind to a protein of interest and using an appropriate linker to attach these molecules to a moiety that binds to the stationary phase.

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Section: Introductionmentioning

confidence: 99%

An Affinity-Based Strategy for the Design of Selective Displacers for the Chromatographic Separation of Proteins

et al. 2008

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“…Ion exchange displacement chromatography has attracted significant attention as a powerful technique for the purification of therapeutic biomolecules because of the high product concentration, purity, and yield that this technique can produce at high column loadings. [1][2][3] While conventional high molecular weight displacers (MW > 2000) such as polyelectrolytes were used in ion exchange systems, 4 it has also been demonstrated that low molecular weight displacers (MW < 2000) can be effective displacers for bioproduct purification. 5 Low MW displacers have significant operational advantages compared to large polyelectrolyte displacers.…”

Section: Introductionmentioning

confidence: 99%

Effect of Displacer Chemistry on Displacer Efficacy for a Sugar-Based Anion Exchange Displacer Library

Liu

Park

Moore

et al. 2006

Ind. Eng. Chem. Res.

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Derivatives (Scheme1). TH(PSNa) 8 was synthesized by the following procedure. To a suspension of NaH (95%, 1.76 g, 70.1 mmols) in anhydrous DMSO was added dropwise trehalose (2.5 g, 7.3 mmols) through a dropping funnel under nitrogen with magnetic stirring. The reaction mixture was heated at 50 o C for 2 hr. Into the suspension 1,3-propansultone (10.0 g, 82.0 mmols) was added dropwise. The reaction mixture was heated at 50 o C for 12 hr. and 80 o C for 12 hr. The mixture was cooled to room temperature and filtered through a fritted filter to remove any precipitate. An ethanol/water mixture (9/1) was added slowly with stirring to cause precipitation. The solid was collected, washed with a mixture of ethanol/water (9/1), and dried at 50 o C under vacuum. The product was purified further by precipitation from DMF/Water (6/1), washing with DMF, and drying under vacuum. Finally, the product was stirred in boiling methanol for 1hr., filtered and dried under vacuum at 60 o C to give 5.9 g of the desired product in 54% yield. 1 H NMR (500MHz, DMSO-d 6 ) δ 4.

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“…Ion-exchange displacement chromatography has attracted significant attention as a powerful technique for the purification of biomolecules in biotherapeutic downstream processes mainly due to the high product concentration, purity, and yield that this technique can produce at high column loadings (1)(2)(3)(4). While conventionally high molecular weight displacers (MW .…”

Section: Introductionmentioning

confidence: 99%

High Throughput Determination and QSER Modeling of Displacer DC‐50 Values for Ion Exchange Systems

Liu

Yang

Ladiwala

et al. 2006

Separation Science and Technology

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In this paper, the displacer concentration required to displace 50% of proteins bound in batch adsorption systems, DC-50, was employed as a means of ranking high-affinity, low molecular weight displacers for ion-exchange systems. A relatively large data set of cationic displacers with varying chemistries were evaluated with two proteins on two strong cation exchange resins in parallel batch screening experiments. Using this methodology, a significant number of high affinity displacers were identified that could displace both proteins at relatively low concentrations. In addition, the DC-50 data was used in concert with molecular structural information of the displacers to produce predictive quantitative structure-efficacy relationship (QSER) models based on a support vector machine (SVM) regression approach. The resulting models were well correlated and the predictive power of these models was demonstrated. Examination of the features selected in these models provided insight into the factors influencing displacer efficacy in cation exchange systems. These results demonstrate the utility of a combined DC-50/ QSER approach to identify and design high-affinity displacers for ion-exchange displacement chromatography.

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Semi-preparative high-performance reversed-phase displacement chromatography of insulins

Cited by 42 publications

References 16 publications

An Affinity-Based Strategy for the Design of Selective Displacers for the Chromatographic Separation of Proteins

An Affinity-Based Strategy for the Design of Selective Displacers for the Chromatographic Separation of Proteins

Effect of Displacer Chemistry on Displacer Efficacy for a Sugar-Based Anion Exchange Displacer Library

High Throughput Determination and QSER Modeling of Displacer DC‐50 Values for Ion Exchange Systems

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