Displacement chromatography of biomolecules

Subramanian, Guhan; Phillips, Michael W.; Cramer, Steven M.

doi:10.1016/s0021-9673(01)83845-0

Cited by 63 publications

(11 citation statements)

References 9 publications

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“…[1][2][3][4][5] Recent reports have demonstrated the efficacy of displacement chromatography for the purification of proteins from industrial process streams. 6,7 Conventionally, large polyelectrolytes have been employed as displacers for ion-exchange systems.…”

Section: Introductionmentioning

confidence: 99%

Synthesis and Characterization of High-Affinity, Low Molecular Weight Displacers for Cation-Exchange Chromatography

Shukla

Bae

Moore

et al. 1998

Ind. Eng. Chem. Res.

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This paper describes the synthesis and characterization of novel low molecular weight displacers for protein purification in cation-exchange systems. A series of displacers based on the pentaerythritol geometry are examined to identify high-affinity, low molecular weight displacers. The steric mass action (SMA) model is employed to evaluate the relative affinities of these displacers. A linear gradient method is described for the measurement of SMA model parameters of molecules. This work demonstrates that, in addition to the number of charges on a displacer molecule, the presence of aromatic moieties can have a profound effect on affinity. In particular, the placement of aromatic groups near the surface of a molecule is shown to result in significant increases in displacer efficacy. Finally, the efficacy of these displacers is examined in displacement experiments using a model protein.These results indicate that nonspecific interactions can play a major role in determining the affinity of a molecule for ion-exchange systems.

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Section: Introductionmentioning

confidence: 99%

Synthesis and Characterization of High-Affinity, Low Molecular Weight Displacers for Cation-Exchange Chromatography

Shukla

Bae

Moore

et al. 1998

Ind. Eng. Chem. Res.

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“…Recent reviews have discussed the trinsically nonlinear mode, has several advantages fundamentals and practice of displacement chromaover the elution mode in process scale separations tography [17][18][19]. [2][3][4][5][6][7][8][9][10][11][12][13][14][15][16]. It may offer the use of larger sample sizes, Displacement chromatography has also been recognized as a method that is especially suited to the separation of closely related substances that differ *Corresponding author.…”

Section: Introductionmentioning

confidence: 99%

“…For displacement chromatography to have utility for biopharmaceutical downstream processing, 2.3.1. Displacement of isomers mixture it must be able to separate some rather difficult ''real Both the displacement separations of the samples world'' mixtures [5,11]. It was reported that by different displacers and the analysis of the rHuBDNF was purified from impurities that have fractions collected during displacement process were extremely similar chromatographic behavior in a all carried out on the same column.…”

Section: Introductionmentioning

confidence: 99%

Displacement chromatography of isomers and therapeutic compounds

Huang

2002

Journal of Chromatography A

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Displacement chromatography was successfully used to separate a binary isomer mixture, epirubicin and doxorubicin, on Kromasil KR100-10 C 25034.6 mm I.D. (10 mm) column. Displacement parameters such as the types and the 18 concentrations of displacer, the composition and the flow rate of the mobile phase were critically examined in this study. The displacer employed was 30 mg / ml benzethonium chloride. Loading of feed at lower initial organic level of mobile phase coupled with displacement at higher organic level was found to give efficient separation. A 30-mg amount of binary isomer mixture was separated on an analytical column. The purification of epirubicin from the closely related impurities present in raw product solution by displacement chromatography was also investigated. The purity of epirubicin required was greater than 99% with a recovery of 60%. The results have indicated that this process made good use of the high feed load, low solvent costs, and high resolution characteristics of displacement chromatography and offered the chromatographic engineer a powerful tool for the preparative purification of therapeutic compounds.

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“…While proteins have also been successfully displaced on metal affinity (Kim and Cramer, 1991;Vunnum et al, 1996) and hydroxyapatite chromatographic supports (Vogt and Freitag, 1997), to date, displacement of proteins has been largely confined to ion-exchange systems (Gerstner et al, 1995;Jayaraman et al, Kundu et al, 1995;Liao et al, 1987;Peterson, 1978;Shukla et al, 1998bShukla et al, , 1998cSubramanian et al, 1988) The displacement of proteins using higher affinity proteins as displacers has been described in HIC systems (Antia et al, 1995). In that work, the authors employed bovine serum albumin (BSA) as a displacer for RNAse A and lysozyme on a butyl HIC column, and ␣-chymotrypsinogen A as a displacer for BSA and lysozyme on a micropellicular HIC resin.…”

Section: Introductionmentioning

confidence: 99%

Hydrophobic displacement chromatography of proteins

Shukla

Sunasara

Rupp

et al. 2000

Biotechnol. Bioeng.

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Displacement chromatography of proteins was successfully carried out in both hydrophobic interaction and reversed‐phase chromatographic systems using low‐molecular weight displacers. The displacers employed for hydrophobic displacement chromatography were water soluble, charged molecules containing several short alkyl and/or aryl groups. Spectroscopy was employed to verify the absence of structural changes to the proteins displaced on these hydrophobic supports. Displacement chromatography on a reversed‐phase material was employed to purify a growth factor protein from its closely related variants, demonstrating the high resolutions that can be achieved by hydrophobic displacement chromatography. This process combines the high‐resolution/high‐throughput characteristics of displacement chromatography with the unique selectivity of these hydrophobic supports and offers the chromatographic engineer a powerful tool for the preparative purification of proteins. © 2000 John Wiley & Sons, Inc. Biotechnol Bioeng 68: 672–680, 2000.

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Displacement chromatography of biomolecules

Cited by 63 publications

References 9 publications

Synthesis and Characterization of High-Affinity, Low Molecular Weight Displacers for Cation-Exchange Chromatography

Synthesis and Characterization of High-Affinity, Low Molecular Weight Displacers for Cation-Exchange Chromatography

Displacement chromatography of isomers and therapeutic compounds

Hydrophobic displacement chromatography of proteins

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