Presentation Information
[4DSP-06-KL]Separation of biologic macromolecules and nanoparticles via convection based chromatographic supports
○ALES PODGORNIK1 (1. FACULTY OF CHEMISTRY AND CHEMICAL TECHNOLOGY, UNIVERSITY OF LJUBLJANA (Slovenia))
Keywords:
convective chromatographic media,monoliths,proteins,DNA,viruses
Convective chromatographic supports are a unique class of stationary phases, specifically designed for the efficient isolation and analysis of large macromolecules and biological nano-assemblies. Unlike membranes, which are limited by their thickness, monoliths can be cast in virtually any geometry or volume—ranging from microliter-scale capillaries to multi-liter cylindrical columns. In contrast to conventional beds composed of spherical particles, monoliths feature diverse microstructures that significantly influence their hydrodynamic and chromatographic performance.
The defining characteristic of monoliths is their continuous, single-piece porous structure. Because the entire porosity consists of interconnected channels, mass transport occurs via convection rather than diffusion. Consequently, the slow diffusivity of large biomolecules is no longer a bottleneck, as they are transported by the mobile phase stream, enabling rapid exchange with the stationary phase. As a result, both separation resolution and dynamic binding capacity remain independent of the flow rate. Furthermore, the high open porosity leads to a low pressure drop even at elevated flow rates, governed by the specific pore size and microstructural topology. However, this high porosity also impacts mechanical stability, a factor that must be carefully addressed during the synthesis of such phases.
In this work, we characterize monoliths with varying microstructures and evaluate their impact on mechanical, hydrodynamic, and chromatographic performance. The latter is demonstrated through the separation of various biological macromolecules and nanoparticles, including proteins, plasmid DNA, and viruses.
The defining characteristic of monoliths is their continuous, single-piece porous structure. Because the entire porosity consists of interconnected channels, mass transport occurs via convection rather than diffusion. Consequently, the slow diffusivity of large biomolecules is no longer a bottleneck, as they are transported by the mobile phase stream, enabling rapid exchange with the stationary phase. As a result, both separation resolution and dynamic binding capacity remain independent of the flow rate. Furthermore, the high open porosity leads to a low pressure drop even at elevated flow rates, governed by the specific pore size and microstructural topology. However, this high porosity also impacts mechanical stability, a factor that must be carefully addressed during the synthesis of such phases.
In this work, we characterize monoliths with varying microstructures and evaluate their impact on mechanical, hydrodynamic, and chromatographic performance. The latter is demonstrated through the separation of various biological macromolecules and nanoparticles, including proteins, plasmid DNA, and viruses.
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