High-performance liquid chromatography of amino acids, peptides and proteins CXXVI. Modelling of protein adsorption with non-porous and porous particles in a finite bath

Mao, Qui-Ming; Stockmann, Regina; Prince, I.G.; Hearn, Milton Thomas William

doi:10.1016/s0021-9673(99)87008-3

Cited by 30 publications

(6 citation statements)

References 22 publications

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“…The particulate media with pore diameter less than 30 nm behave like a non-porous medium [40,41] when loaded with plasmids. The DBC was independent of flow rate and feed concentration, but lower by one order of magnitude compared to the monolith.…”

Section: Resultsmentioning

confidence: 99%

Mass transfer characteristics of plasmids in monoliths

Zöchling

Hahn

Ahrer

et al. 2004

J of Separation Science

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The hydrodynamic properties and pore-structure of monoliths based on functionalized poly(glycidyl methacrylate-ethylene dimethacrylate) were characterised by pulse response experiments using different probes representing a wide range of molecular mass. On a small scale, band spreading was found to be caused to the extent of more than 90% by extra-column effects. These monoliths have large channel diameters, providing a suitable chromatography adsorbent for processing of large molecules. Dynamic and static binding capacity for plasmid DNA was investigated. For our model plasmid, consisting of 4.9 kbp, a capacity of 7 mg/mL was observed in comparison to 0.3 mg/mL for a conventional medium designed for protein separation. When plasmids were loaded on the monolith a gradual increase in pressure drop was observed. The channels filled up and the cross-sectional area available for liquid flow decreased. Therefore, a higher pressure drop was observed during elution. This is caused by (i) shrinking of the channels as effect of the high salt concentration, (ii) high viscosity of the mobile phase due to high concentration of plasmids, and (iii) an increase of the hydrodynamic radius of the plasmid with salt concentration from 45 nm at 150 mM to 70 nm at 2 M NaCl, as measured by dynamic light scattering. These types of monoliths are considered to be the preferred adsorbents for plasmid separation.

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Section: Resultsmentioning

confidence: 99%

Mass transfer characteristics of plasmids in monoliths

Zöchling

Hahn

Ahrer

et al. 2004

J of Separation Science

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“…Combining eqs 6, 9, 10, and 12, the differential form of the concentration profile that accounts for the change in the bulk fluid concentration of plasmid is (12) (assuming a Langmuir isotherm): In the last equation, x 1 and x 2 are the roots of the quadratic equation and K D = 1/ K A is the Langmuir dissociation constant.…”

Section: Theorymentioning

confidence: 99%

Studies on the Batch Adsorption of Plasmid DNA onto Anion-Exchange Chromatographic Supports

Ferreira¹,

Cabral²,

Prazeres³

2000

Biotechnol. Prog.

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The adsorption of a supercoiled 4.8 kbp plasmid onto quaternary ammonium anion-exchangers was studied in a finite bath. Equilibrium experiments were performed with pure plasmid, at 25 degrees C, using commercial Q-Sepharose matrices differing in particle diameter (High Performance, 34 microm; Fast Flow, 90 microm; and Big Beads, 200 microm) and a recently commercialized ion-exchanger, Streamline QXL (d(p) = 200 microm) at different salt concentrations (0.5, 0.7, and 1 M NaCl). Plasmid adsorption was found to follow second-order kinetics (Langmuir isotherm) with average association constants K(A) = 0.32+/-0.12 mL microg(-)(1) and K(A) = 0.25+/-0.15 mL microg(-1) at 0.5 and 0.7 M Nacl, respectively. The maximum binding capacities were not dependent on the ionic strength in the range 0.5-0.7 M but decreased with increasing particle diameter, suggesting that adsorption mainly occurs at the surface of the particles. No adsorption was found at 1 M NaCl. A nonporous model was applied to describe the uptake rate of plasmid onto Streamline QXL at 0.5 M NaCl. The overall process rate was controlled by mass transfer in the regions of low relative amounts of adsorbent (initial stages) and kinetically controlled in the later stages of the process for high relative amounts of adsorbent. The forward reaction rate constant (k(1) = 0.09+/-0.01 mL mg(-1) s(-1)) and film mass transfer coefficient (K(f) = (6 +/- 2) x 10(-4) cm s(-1)) were calculated. Simulations were performed to study the effect of the relative amount of adsorbent on the overall process rate, yield, and media capacity utilization.

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“…Adsorption and transport processes under the Langmuir isotherm assumptions have been modelled mathematically 3 to better understand the phenomena involved in capillary electrochromatography. 4,5 Several models exist for protein mass transfer, 6 for their adsorption on ion exchange particles [7][8][9][10] and on sorbent matrices. 11,12 Mathematical models have also been used to describe the adsorption kinetics of proteins 13 and polyelectrolytes 14 on planar surfaces.…”

Section: Introductionmentioning

confidence: 99%

Protein adsorption in static microsystems: effect of the surface to volume ratio

2005

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A numerical model for the adsorption kinetics of proteins on the walls of a microchannel has been developed using the finite element method (FEM) to address the coupling with diffusion phenomena in the restricted microchannel volume. Time evolutions of the concentration of one species are given, both in solution and on the microchannel walls. The model illustrates the adsorption limitation sometimes observed when the microdimensions of these systems induce a global depletion of the bulk solution. A new non-dimensional parameter is introduced to predict the final value of the coverage of any microsystem under static adsorption. A working curve and a criteria (h/KC max . 10) are provided in order to choose, for given adsorption characteristics, the value of the volume-to-surface ratio (i.e. the channel height h) avoiding depletion effects on the coverage (relative coverage greater than 90% of the theoretical one). Simulations were compared with confocal microscopy measurements of IgG antibody adsorption on the walls of a PET microchannel. The fit of the model to the experimental data show that the adsorption is under apparent kinetic control.

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High-performance liquid chromatography of amino acids, peptides and proteins CXXVI. Modelling of protein adsorption with non-porous and porous particles in a finite bath

Cited by 30 publications

References 22 publications

Mass transfer characteristics of plasmids in monoliths

Mass transfer characteristics of plasmids in monoliths

Studies on the Batch Adsorption of Plasmid DNA onto Anion-Exchange Chromatographic Supports

Protein adsorption in static microsystems: effect of the surface to volume ratio

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