Design and preparation of novel particulate and continuous polymeric macroporous media for the separation of biological and synthetic molecules

Abstract Novel approaches to macroporous polymeric separation media with enhanced pore size distributions, controlled surface chemistries, and excellent separation properties have been explored. In the first of these approaches, size‐monodisperse spherical polymeric particles containing both large and small pores are prepared with the use of a variety of polymeric porogens. The choice of polymeric porogen as well as its concentration greatly affects the pore size and pore size distribution of the final beads. The polymer beads obtained by this technique have been tested in size exclusion chromatography and in the separation of proteins in reversed‐phase mode. Techniques that allow the specific functionalization of pores according to their sizes have been developed. Thus, the surfaces of the large pores of the monodisperse particles can be made hydrophilic or chiral while the smaller pores remain hydrophobic or achiral. Similarly, a novel polymeric separation medium based on a continuous polymer rod incorporating macropores has been prepared by an in situ polymerization within the confines of the tube of a chromatographic column. For example, non‐particulate continuous polymer rods with flow‐through pores as well as separation pores have been prepared from styrene and divinylbenzene and used in the very fast (30 s) separation of proteins by reversed‐phase chromatography. Alternatively, continuous rods prepared from glycidyl methacrylate and ethylene dimethacrylate can be transformed into materials that are suitable for ion exchange or other chromatographic modes. A characteristic of the continuous polymeric rod media is their permeability which allows the mobile phase to flow through the medium without causing a high back‐pressure. The continuous rod‐shaped media can also be modified in pore size‐selective fashion to introduce different chemistries in the large and the small pores. These new media are advantageous as they are extremely easy to prepare, durable, require no packing, and can be used for a variety of separations.