Monoliths for fast bioseparation and bioconversion and their applications in biotechnology

Abstract

Monoliths have consolidated their position in bioseparation. More than 200 different applications have been reported in the past two decades and their advantages compared to conventional chromatography demonstrated. These include the high mass transfer efficiency due to the convective flow enabled by the macroporous character of the matrix. Recently plasmid DNA and viruses were separated with high efficiency and cryogels and monolithic superporous agarose were developed for capture of proteins from crude homogenates and separation of microorganisms or lymphocytes. Currently four companies manufacture monoliths mainly for analytical applications although monoliths with a volume of 0.8 liter are commercially available and 8 L are available as prototypes. A book entitled "Monolithic materials: preparation, properties and applications" was published in 2003 and became standard reference of the status of this area. This review focuses on the progress in monoliths that goes beyond the scope of this reference book. Less progress has been made in the field of bioconversions in spite of the fact that monolithic supports exhibit better performance than beads in enzymatic processing of macromolecules. It appears that the scientific community has not yet realized that supports for these applications are readily available. In addition, monoliths will further substantially advance bioseparations of both small and large molecules in the future.

Adsorption of plasmid DNA on anion exchange chromatography media

“…Regarding also the effective diffusivity obtained from the pore diffusion model, it seems that mass transfer is faster than predicted and diffusion limitation is negligible. According to theory there is no mass transfer limitation in the large channels of monolithic supports, which are governed by convective flow [8,16,46,47]. Results of CIM DEAE (Fig.…”

Rapid production of a plasmid DNA encoding a malaria vaccine candidate via amino‐functionalized poly(GMA‐co‐EDMA) monolith

“…The most essential feature of monolithic supports is that all of the mobile phase is forced to flow through the large pores of the monolith, thereby allowing the penetration of large pDNA molecules to the internal surface area, but at reduced pressure drops, even at high-flow rates. [15][16][17][18] This convective mass transfer enhances plasmid retention, binding capacity, purification yield or productivity and the economics of the purification process. Here we report a cost-effective, nontoxic and scalable technique for the rapid isolation of a pDNA encoding PyMSP4/5, a homologue of P. falciparum merozoite surface protein 4 (MSP4) and 5 (MSP5) in rodent malaria species P. yoelii.…”

Less common applications of monoliths

“…This activity led to acceptance of monolithic columns in the large family of chromatographic stationary phases. Their applications in a variety of liquid chromatographic modes including high-performance liquid chromatography (HPLC) and capillary electrochromatography (CEC) has recently been described in several reviews [12][13][14][15][16][17][18][19][20][21][22][23][24][25] and books [11,26]. However, the less common applications of monolithic materials that include supports for solid phase and combinatorial synthesis [27][28][29], scavengers [30,31], carriers for immobilization of enzymes [32][33][34], static mixers [35], thermally responsive gates and valves [36][37][38], as well as solid phase extractors and pre-concentrators [39] are escaping the awareness of the scientific community.…”