Affinity chromatography of proteins on monolithic columns

At present, monolithic stationary phases, because of their morphology, are widely used for development and realization of fast dynamic and static processes based on mass transition between liquid and solid phases. These are liquid chromatography, solid phase synthesis, microarrays, flow-through enzyme reactors, etc. High-performance liquid chromatography on monoliths, including bioaffinity mode, represents a unique technique appropriate for fast and efficient separation of biological (macro)molecules of different sizes and shapes (proteins, nucleic acids, peptides), as well as such supramolecular systems as viruses.In this work, the examples of application of commercially available macroporous monoliths for modern affinity processing are presented. In particular, the original methods developed for efficient isolation and fractionation of monospecific antibodies from rabbit blood sera, the possibility of simultaneous affinity separation of protein G and serum albumin from human serum, the isolation of recombinant products, such as protein G and tissue plasminogen activator from E. coli cell lysate and Chinese Hamster Ovary cell culture supernatant, respectively, are described in detail. The suggested and realized multifunctional fractionation of polyclonal pools of antibodies by combination of several short monolithic columns (disks) with different affinity functionalities stacked in the same cartridge represents an original and practically valuable method that can be used in biotechnology.