Premium
Methacrylate‐based short monolithic columns: Enabling tools for rapid and efficient analyses of biomolecules and nanoparticles
Author(s) -
Barut Miloš,
Podgornik Aleš,
Urbas Lidija,
Gabor Boštjan,
Brne Peter,
Vidič Jana,
Plevčak Sašo,
Štrancar Aleš
Publication year - 2008
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200800189
Subject(s) - monolith , biomolecule , monolithic hplc column , bioprocess , methacrylate , computer science , polymer , process (computing) , column (typography) , materials science , nanotechnology , chemistry , chromatography , stationary phase , chemical engineering , polymerization , catalysis , engineering , telecommunications , biochemistry , operating system , frame (networking) , composite material
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.