Rate parameters and gradient correlations for gradient‐elution chromatography

Luo, Robert G.; Hsu, Jong-Ping

doi:10.1002/aic.690430219

Cited by 9 publications

(14 citation statements)

References 38 publications

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“…The good separation achieved indicated that the packed beads in the shell side were working well as an anion exchanger; further, the packed bed column with the fibers was an effective axial flow chromatographic column. The peak shape for β‐LG is quite similar to that obtained by Luo and Hsu (23) in packed bed chromatography with DEAE‐Sepharose beads in a bed that was 1.5 cm in diameter and 22.0 cm in length. The elution of β‐LG could have been initiated earlier by an earlier introduction of 0.5 M NaCl.…”

Section: Resultssupporting

confidence: 83%

“…Correspondingly, the model proteins employed will demonstrate the isolation and purification capabilities of the integrated device and process. These model proteins are widely used by other researchers as well (23, 24). Industrial application of the integrated process and device to specific product protein needs to be developed on a case by case basis.…”

Section: Introductionmentioning

confidence: 99%

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An Integrated Process for Biomolecule Isolation and Purification

1999

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Biomolecule isolation and purification from a fermentation broth usually involve centrifugation, filtration, adsorption, and chromatography steps. Each step contributes to the product cost and product loss. In this research, a cyclic process integrating commercially available ultrafiltration membranes and chromatographic resin beads was developed to achieve the same goal in one device. The device consisted of ion exchange beads on the shell side of a hollow fiber ultrafiltration module. Loading of proteins on the stationary phase on the shell side was carried out for a period of 5-20 min from the permeate on the shell side produced from tube-side feed in ultrafiltration. The eluent was then introduced either from the shell-side inlet or tube-side inlet; the chromatographic fractions were collected from the shell-side outlet. The column was regenerated/washed next to start a new cycle. Systems studied in this cyclic process include the following binary mixtures: myoglobin and beta-lactoglobulin; hemoglobin and bovine serum albumin; and myoglobin and alpha-lactalbumin. Excellent resolutions of the proteins were obtained. A yeast-based cellular suspension containing a mixture of myoglobin and alpha-lactalbumin was also applied to this device. The target proteins were recovered and purified successfully. The cyclic process-based device integrates clarification, concentration, and chromatographic purification of biomolecules and is suitable for both extracellular and intracellular products.

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Section: Resultssupporting

confidence: 83%

Section: Introductionmentioning

confidence: 99%

An Integrated Process for Biomolecule Isolation and Purification

1999

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“…LGA and LGB differ in their primary structure at two amino acid residues, at position 64 and 118, where LGA has aspartic acid at 64 and valine at 118, and LGB has glycine at 64 and alanine at 118, with a total of 162 residues each. They both have a pI in the range of 4.9‐5.1 . The other protein that is analyzed in this work is bovine serum albumin (BSA; Sigma‐Aldrich, St. Louis, MO), which has a similar pI between 4.7 and 5.3 .…”

Section: Methodsmentioning

confidence: 99%

The pH, temperature, and protein structure effect on β‐lactoglobulin A and B separation in anion‐exchange chromatography

Pavlov

Hsu

2018

AIChE Journal

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The effect of pH and temperature on separating a mixture of similar proteins, namely b-lactoglobulin A (LGA) and b-lactoglobulin B (LGB) in anion-exchange chromatography is explored. The proteins carry a slight difference in negative charge at basic pH, providing a separation basis on an Q Sepharose Fast Flow anion-exchange resin. They were separated at different temperatures and pH values, and the separation factor was evaluated. The experimental results were matched to a theoretical model to compute the equilibrium constant K A . The data shows that an increase in temperature and pH leads to an increase in the retention time of the proteins. The results were correlated with the net charge of the molecule for the separation so that the elution can be simulated for any condition that was studied. The tertiary structures of LGA and LGB are analyzed to illustrate the structure effect on the separation.

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“…Thus, (D ieff /u i ) would remain constant despite any change in ionic strength. Luo and Hsu (1997) found that the D ieff for the DEAE-Sepharose CL-6B ion exchanger was 0.235 cm 2 /min (3.917 × 10 −7 m 2 /s). The D ieff /u i value was 7.83 × 10 −4 m for the beads used in their study.…”

Section: Estimation Of Parametersmentioning

confidence: 99%

“…These beads have the same size distribution as that of DEAE-Sepharose CL-6B ion exchanger. Thus, it can be assumed that the ratio of D ieff /u iP in this study would be equal to that of Luo and Hsu (1997)-that is, 7.83 × 10 −4 m; therefore, the following equation can be adopted:…”

Section: Estimation Of Parametersmentioning

confidence: 99%

Protein loading, elution, and resolution behavior in a novel device that integrates ultrafiltration and chromatographic separation

Dai

Majumdar

Luo

et al. 2003

Biotech & Bioengineering

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Abstract:Hollow fiber membranes and chromatographic resin beads are commonly employed in a variety of bioseparation processes. A new class of integrated separation devices is being studied in which the shell side of a hollow fiber device is filled with adsorbents/ chromatographic resin beads. Such devices and the corresponding separation methods integrate feed broth clarification by the microfiltration/ultrafiltration membrane with bioproduct purification by the shell-side resin beads either as an adsorbent or as beads in elution chromatography. A mathematical model has been developed for the prediction of the chromatographic behavior of such an integrated device. Simulations have been done to study the effects of axial dispersion, feed flow rate, water permeation rate, fiber packing density, and void fraction. Numerical solutions were obtained by solving the governing equations. This model can reasonably describe the concentration profiles as well as the breakthrough and elution behaviors in the integrated device.

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Rate parameters and gradient correlations for gradient‐elution chromatography

Cited by 9 publications

References 38 publications

An Integrated Process for Biomolecule Isolation and Purification

An Integrated Process for Biomolecule Isolation and Purification

The pH, temperature, and protein structure effect on β‐lactoglobulin A and B separation in anion‐exchange chromatography

Protein loading, elution, and resolution behavior in a novel device that integrates ultrafiltration and chromatographic separation

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