Effect of Intraparticle Convection on the Chromatography of Biomacromolecules

Abstract

The effect of intraparticle convection in chromatographic columns packed with gigaporous particles (i.e., where dpore/dparticle > 10(-2)) on the band spreading of unretained biomacromolecules is investigated both experimentally and theoretically. A model is developed for the analysis of mass transfer in spherical particles of bidisperse pore structure when both convection and diffusion take place in the larger pores but only diffusion occurs in the smaller pores. The predictions of the model were experimentally verified. It is demonstrated that gigaporous particles have advantages over conventional porous particles (i.e., where dpore/dparticle < 10(-3)) for applications that do not require high resolving power, to bring about fast separation. This is because columns packed with gigaporous particles can be operated at high flow velocities without significant loss of efficiency due to the enhancement of mass transfer by intraparticle convection. The results of the model are used to examine the effectiveness of gigaporous column packings for rapid analytical chromatography and for the concentration and recovery of a dilute solute in a saturation-regeneration cycle utilizing frontal chromatography.

Impact of denaturation with urea on recombinant apolipoprotein A‐IMilano ion‐exchange adsorption: Equilibrium uptake behavior and protein mass transfer kinetics

“…The effective pore diffusivity is related to the free solution diffusivity according to [27]: (10) where τ is the particle tortuosity factor and λ is a hindrance parameter. Porosity and tortuosity are properties of the stationary phase and should remain similar as the ionic strength of the buffers used in breakthrough experiments is essentially equal in all cases.…”

“…For bioprocess applications where large diameter adsorbent beads and high mobile phase velocities are typically utilized, chromatographic efficiency is nearly completely determined by mass transfer kinetics [25]. Accordingly, numerous studies have been devoted to protein mass transfer in chromatography media (e.g., [26][27][28][29][30][31]). However, to our knowledge, the effects of urea on protein-binding capacity, chromatography retention, and intraparticle mass transfer in ion-exchange columns have not been investigated in a systemic fashion.…”

Equilibrium and Kinetics of High Molecular Weight Protein Uptake in Ion Exchange Chromatography

“…In this context, Frey et al [37] introduced a correlation between the effective diffusivity (D e ) and the free aqueous solution diffusivity (D 0 ) as it is indicated in the following equation: (14) where ε p is the particle porosity, τ is the particle tortuosity and λ [6] is the steric hindrance factor. For a chromatographic medium, the ratio ε p /τ is constant.…”

The use of confocal laser scanning microscopy to study the transport of biomacromolecules in a macroporous support

“…Kinetics models are used to predict the parameters that characterize the intraparticle mass transfer mechanisms. Several investigators have approached the prediction of transport rates using the plate height method to account for the combined effects of convection and diffusion (2)(3)(4)(5). Although these approaches have shown to be effective in predicting the effects of transport in particular chromatographic environments, these models present limitations.…”