Protein separation by liquid chromatography using permeable POROS Q/M particles

Rodrigues, Alírio E.; Chenou, C.; Rendueles, Manuel

doi:10.1016/0923-0467(95)03045-x

Cited by 11 publications

(5 citation statements)

References 18 publications

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“…The decreasing slope of the experimental HETP curves as u 0 increases is due to convective enhancement that reduces the overall intraparticle mass transfer resistance. Celbeads‐CM shows typical behavior of macroporous adsorbent 57. The indicated critical velocity, u 0c , for commencement of convective flow in the pores is about 1.5 cm/min and H plateau is about 0.5 cm.…”

Section: Resultsmentioning

confidence: 95%

“…In laminar flow, the pressure drop Δ P across a bed of a length L packed with particles of diameter d p is given by Darcy's law, Δ P / L = η u 0 / B b , where η is the fluid viscosity and u 0 is the superficial velocity. Bed permeability, B b , is obtained from the slope of the plot Δ P / L vs. the bed superficial velocity, u 0 , B b = (ε

_{3}^{b}

_{2}^{p}

/[150 (1 − ε b ) 2 ] and ε b is the bed porosity 57. Figure 5 shows bed pressure drop as a function of u 0 for lysozyme solution.…”

Section: Resultsmentioning

confidence: 99%

“…HETP experiments were performed under nonretaining conditions in elution buffer on celbeads‐CM in packed‐bed as well as EB operation. The HETP as a function of superficial velocity is calculated from the experimental chromatographic peaks by HETP = σ 2 L /μ

_{2}^{1}

, where σ 2 is the peak variance, μ 1 is the first moment of the peak, and L is the column length 57. From the first moment of the peaks, we estimated the total bed porosity, ε t = ε b + (1 − ε b )ε p , and thus the particle porosity is ε p = 0.58.…”

Section: Resultsmentioning

confidence: 99%

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Characterization of superporous cellulose matrix for high‐throughput adsorptive purification of lysozyme

Kale

Lali

2011

Biotechnology Progress

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Protein purification essentially requires macroporous adsorbents matrices, which can provide high efficiency in packed bed and expanded bed (EB) even at high flow rates on account of reduced pore diffusion resistance resulting from finite intraparticle flow in the superpores. Rigid spherical superporous adsorbent beads with high carboxyl group density were prepared by crosslinking of cellulose. The matrix (diameter: 100-300 μm, mean pore size: 1-3 μm, pore volume: 57-59%, and bulk density: ∼1,438 kg/m3) could be used in packed bed as well as EB for purification of various biomolecules. Attempts were made to use indigenously developed rigid, superporous crosslinked cellulose adsorbent for high-throughput purification of lysozyme from chicken egg white's extract. A typical adsorption isotherm for lysozyme in crude was well correlated with the Langmuir isotherm model. Two maxima of binding capacity on celbeads bearing carboxymethyl (celbeads-CM) group for lysozyme were observed at pH 4.5 and 7.5. Uptake kinetics showed that the diffusivity of lysozyme was 100 times higher than conventional matrices. Such superporous matrix can be used for high-throughput purification of proteins from crude feedstocks and is reflected in leveling off of height equivalent to theoretical plate vs. flow curve after threshold velocity. Optimization of binding and elution conditions resulted in overall purification of lysozyme in a high yield and purity of 98.22 and 98.8%, respectively, with purification factor of 51.54 in a single step. The overall productivity (14.21 kg/m3 h) and specific activity (2.2×10(5) U/mg) were higher than that obtained with traditional particulate resins.

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Section: Resultsmentioning

confidence: 95%

_{3}^{b}

_{2}^{p}

/[150 (1 − ε b ) 2 ] and ε b is the bed porosity 57. Figure 5 shows bed pressure drop as a function of u 0 for lysozyme solution.…”

Section: Resultsmentioning

confidence: 99%

_{2}^{1}

Section: Resultsmentioning

confidence: 99%

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Characterization of superporous cellulose matrix for high‐throughput adsorptive purification of lysozyme

Kale

Lali

2011

Biotechnology Progress

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“…Finally, it is noted that the pore and solid diffusion models, Eqs. (2) and (3), are limiting cases of the general parallel diffusion model Eq. (4).…”

Section: Parallel Diffusionmentioning

confidence: 99%

The Influence of Retention on the Plate Height in Ion‐Exchange Chromatography

Hansen¹,

Mollerup²

2005

Separation Science and Technology

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The plate heights for the amino acid tyrosine (anion exchange) and the polypeptide aprotinin (cation exchange) were determined on a porous media (Resource 15) and a gel filled media (HyperD 20) at salt concentrations ranging from weak to strong retention. At a constant velocity, measurements showed that the plate height increase with increasing retention, went through a maximum, and finally, decreased as the retention increased, i.e., when the salt concentration was lowered further. The band broadening of a chromatographic peak in the column was caused by the axial dispersion and mass transfer. In this article, the rate of mass transfer in the particles is described by three different rate mechanisms, pore diffusion, solid diffusion, and parallel diffusion. The van Deemter equation was used to model the data to determine the mass-transfer properties. The development of the plate height with increasing retention revealed a characteristic behavior for each rate mechanism. In the pore diffusion model, the plate height increased toward a constant value at strong retention, while the plate height in the solid diffusion model decreased, approaching a constant value at strong retention. In the parallel diffusion model, both pore and solid diffusion took place. Therefore, the parallel diffusion model coincides with the pore diffusion model at weak retention and with the solid diffusion model at strong retention, while a maximum is reached at intermediate retention, resulting in a bell-shaped curve. This behavior corresponds to the observed variation of the plate height at constant velocity. Neither the pore nor the solid diffusion model can describe the experimental data while a satisfactory fit was obtained using the parallel diffusion model.

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“…Significant effects of intraparticle mass transfer on separation result in single chromatographic column have been realized by many researchers [1][2][3][4][5][6][7][8][9][10]. But the effects of intraparticle mass transfer on simulated moving bed (SMB) process have seldom been reported.…”

Section: Introductionmentioning

confidence: 99%

Comprehension of Effects of Intraparticle Effective Diffusivity on Simulated Moving Bed

Gao¹

2017

J Chromatogr Sep Tech

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Protein separation by liquid chromatography using permeable POROS Q/M particles

Cited by 11 publications

References 18 publications

Characterization of superporous cellulose matrix for high‐throughput adsorptive purification of lysozyme

Characterization of superporous cellulose matrix for high‐throughput adsorptive purification of lysozyme

The Influence of Retention on the Plate Height in Ion‐Exchange Chromatography

Comprehension of Effects of Intraparticle Effective Diffusivity on Simulated Moving Bed

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