Hydrophobic interaction chromatography of proteins

Machold, Christine; Deinhofer, Karin; Hahn, Rainer; Jungbauer, Alois

doi:10.1016/s0021-9673(02)01077-4

Cited by 81 publications

(50 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Some initial attempts to introduce 'low throughput' screening of chromatographic resins for a given task can be found in the literature [6][7][8]. An increase in sample throughput and thus data generation can be achieved by automation, parallelization and miniaturization.…”

Section: Introductionmentioning

confidence: 99%

High Throughput Screening for the Design and Optimization of Chromatographic Processes – Miniaturization, Automation and Parallelization of Breakthrough and Elution Studies

et al. 2008

View full text Add to dashboard Cite

This study evaluates the applicability of the liquid handling workstation Tecan Evo Freedom 200 and 200 lL Media Scout RoboColumns to dynamic chromatographic operations such as frontal analysis (breakthrough) and elution experiments in a high throughput screening mode. Breakthrough experiments were conducted using BSA as a model protein and the stationary phases Poros 50 D, Q Ceramic HyperD and DEAE Sepharose FF. The obtained dynamic capacities at 10 and 50 % breakthrough matched well with reference data. Elution experiments were performed applying three protein mixtures: a) lipolase and BSA, b) human growth hormone and a process-related impurity, and c) an insulin analogue and a process-related impurity. The resins used resembled a variety of different ion exchange resins. In all cases, the resulting elution curves matched well with reference measurements performed on an ÄKTA explorer system at 1 mL or 2 mL scale.

show abstract

Section: Introductionmentioning

confidence: 99%

High Throughput Screening for the Design and Optimization of Chromatographic Processes – Miniaturization, Automation and Parallelization of Breakthrough and Elution Studies

et al. 2008

View full text Add to dashboard Cite

show abstract

“…Hydrophobic interaction chromatography (HIC) has been shown to have significant utility for the separation of proteins from complex mixtures (Diogo et al, 2000(Diogo et al, , 2001Husi and Walkinshaw, 1999;Machold et al, 2003;O'Connor et al, 2000;Pomazal et al, 2002;Sunasara et al, 2003). HIC stationary phases are manufactured by attaching relatively weak hydrophobic functional groups to an agarose or polymer backbone .…”

Section: Introductionmentioning

confidence: 99%

Evaluation of selectivity changes in HIC systems using a preferential interaction based analysis

Xia

Nagrath

Garde

et al. 2004

Biotech & Bioengineering

View full text Add to dashboard Cite

It is well established that salt enhances the interaction between solutes (e.g., proteins, displacers) and the weak hydrophobic ligands in hydrophobic interaction chromatography (HIC) and that various salts (e.g., kosmotropes, chaotropes, and neutral) have different effects on protein retention. In this article, the solute affinity in kosmotropic, chaotropic, and neutral mobile phases are compared and the selectivity of solutes in the presence of these salts is examined. Since solute binding in HIC systems is driven by the release of water molecules, the total number of released water molecules in the presence of various types of salts was calculated using the preferential interaction theory. Chromatographic retention times and selectivity reversals of both proteins and displacers were found to be consistent with the total number of released water molecules. Finally, the solute surface hydrophobicity was also found to have a significant effect on its retention in HIC systems. B 2004 Wiley Periodicals, Inc.

show abstract

“…Se ha logrado establecer que las principales variables de proceso son el tipo y la concentración máxima de sal utilizada para formar el gradiente de elución (Melander et al, 1989;Fausnaugh y Regnier, 1986) y el tipo de matriz cromatográfica (Machold et al, 2002), mientras que de las propiedades de las proteínas, la más importante sería la hidrofobicidad (Queiroz et al, 2001;Lienqueo et al, 2002).…”

Section: Introductionunclassified

Modelación Fenomenológica de Cromatografía de Interacción Hidrofóbica

Mahn

2009

Inf. tecnol.

View full text Add to dashboard Cite

ResumenEl objetivo de este trabajo fue obtener un modelo fenomenológico que describa el perfil de elusión, en cromatografía de interacción hidrofóbica de una proteína inyectada como pulso. Se adaptó el modelo de platos, el que fue resuelto numéricamente, obteniéndose un buen ajuste a los datos experimentales. El modelo de platos permitió realizar simulaciones del proceso bajo distintas condiciones de operación, como velocidad de flujo, gradiente de elución y volumen de muestra inyectada. Se concluye que es posible aplicar la teoría de platos para describir perfiles de elución de proteínas en cromatografía de interacción hidrofóbica. El modelo obtenido podría ser utilizado para optimizar la separación de una mezcla de proteínas. Palabras clave: cromatografía, interacción hidrofóbica, purificación de proteínas, separación diferencial Phenomenological Modelling of Hydrophobic Interaction Chromatography AbstractThe aim of this work was to obtain a phenomenological model to describe the elution profile in hydrophobic interaction chromatography of a protein injected as a pulse. The plate model, which was solved numerically, was used, and good fit to experimental data was found. The plate model allowed simulating the chromatographic process under different operating conditions, such as flow rate, elution gradient and sample volume. In is concluded that it is possible to apply the plate theory to describe elution profiles of proteins in hydrophobic interaction chromatography. The proposed model could be used to optimize the separation of protein mixtures.

show abstract

Hydrophobic interaction chromatography of proteins

Cited by 81 publications

References 24 publications

High Throughput Screening for the Design and Optimization of Chromatographic Processes – Miniaturization, Automation and Parallelization of Breakthrough and Elution Studies

High Throughput Screening for the Design and Optimization of Chromatographic Processes – Miniaturization, Automation and Parallelization of Breakthrough and Elution Studies

Evaluation of selectivity changes in HIC systems using a preferential interaction based analysis

Modelación Fenomenológica de Cromatografía de Interacción Hidrofóbica

Contact Info

Product

Resources

About