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Macroporous Polymer Monoliths for Affinity Chromatography and Solid-Phase Enzyme Processing

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Book cover Protein Downstream Processing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2178))

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Abstract

Nowadays, monolithic stationary phases, because of their special morphology and enormous permeability, are widely used for the development and realization of fast dynamic and static processes based on the 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 the bioaffinity mode, represents 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 the edited chapter, the examples of the 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, respectively, are described in detail. The suggested and realized multifunctional fractionation of polyclonal pools of antibodies by the combination of several short monolithic columns (disks) with different affinity functionalities stacked in the same cartridge represents the original and practically valuable method that can be used in biotechnology. In addition, macroporous monoliths were adapted to the immobilization of such different enzymes as polynucleotide phosphorylase, ribonuclease A, α-chymotrypsin, chitinolytic biocatalysts, β-xylosidase, and β-xylanase. The possibility of use of immobilized enzyme reactors based on monoliths for different purposes is demonstrated.

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Acknowledgments

The authors are very grateful to BIA Separations for long-term fruitful cooperation.

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Authors and Affiliations

  1. Institute of Macromolecular Compounds, Russian Academy of Sciences, St. Petersburg, Russia

    E. G. Korzhikova-Vlakh & G. A. Platonova

  2. Institute of Chemistry, Saint-Petersburg State University, St. Petersburg, Russia

    T. B. Tennikova

Authors
  1. E. G. Korzhikova-Vlakh
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  2. G. A. Platonova
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  3. T. B. Tennikova
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Editors and Affiliations

  1. Laboratory of Enzyme Technology, Department of Biotechnology, School of Applied Biology and Biotechnology, Agricultural University of Athens, Athens, Greece

    Nikolaos E. Labrou

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Korzhikova-Vlakh, E.G., Platonova, G.A., Tennikova, T.B. (2021). Macroporous Polymer Monoliths for Affinity Chromatography and Solid-Phase Enzyme Processing. In: Labrou, N.E. (eds) Protein Downstream Processing. Methods in Molecular Biology, vol 2178. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0775-6_18

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  • DOI: https://doi.org/10.1007/978-1-0716-0775-6_18

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0774-9

  • Online ISBN: 978-1-0716-0775-6

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