Methacrylate-based short monolithic columns: Enabling tools for rapid and efficient analyses of biomolecules and nanoparticles

• Published in: Journal of separation science 2008-06, Vol.31 (11), p.1867-1880
• Main Authors: Barut, Miloš, Podgornik, Aleš, Urbas, Lidija, Gabor, Boštjan, Brne, Peter, Vidič, Jana, Plevčak, Sašo, Štrancar, Aleš
• Format: Article
• Language: English
• Subjects: Analytical biochemistry: general aspects, technics, instrumentation; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Chemistry Techniques, Analytical - instrumentation; Chromatography, High Pressure Liquid - instrumentation; Chromatography, Ion Exchange - instrumentation; CIM; DNA; DNA - analysis; DNA - isolation & purification; Fundamental and applied biological sciences. Psychology; Humans; Methacrylates; Monoliths; Nanoparticles - analysis; Proteins; Proteins - analysis; Proteins - isolation & purification; Short monolithic columns
• ISSN: 1615-9306; 1615-9314
• DOI: 10.1002/jssc.200800189

This review describes the novel chromatography stationary phase - a porous monolithic methacrylate-based polymer - in terms of the design of the columns and some of the features that make these columns attractive for the purification of large biomolecules. We first start with a brief summary of the characteristics of these large molecules (more precisely large proteins like immunoglobulins G and M, plasmid deoxyribonucleic acid (DNA), and viral particles), and a list of some of the problems that were encountered during the development of efficient purification processes. We then briefly describe the structure of the methacrylate-based monolith and emphasize the features which make them more than suitable for dealing with large entities. The highly efficient structure on a small scale can be transferred to a large scale without the need of making column modifications, and the various approaches of how this is accomplished are briefly presented in this paper. This is followed by presenting some of the examples from the bioprocess development schemes, where the implementation of the methacrylate-based monolithic columns has resulted in a very efficient and productive process. Following this, we move back to the analytical scale and demonstrate the efficiency of the monolithic column - where the mass transfer between the stationary and mobile phase is greatly enhanced - for the in-process and final control of the new therapeutics. The combination of an efficient structure and the appropriate hardware results in separations of proteins with residence time less than 0.1 s.