TY - CHAP
T1 - Macroporous polymer monoliths for affinity chromatography and solid-phase enzyme processing
AU - Korzhikova-Vlakh, E. G.
AU - Platonova, G. A.
AU - Tennikova, T. B.
N1 - 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
PY - 2021
Y1 - 2021
N2 - 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.
AB - 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.
KW - Affinity chromatography
KW - Conjoint chromatography
KW - Enzyme reactors
KW - Enzymes
KW - Fractionation
KW - Monoliths
KW - Protein and peptide immobilization
KW - Proteins
UR - http://www.scopus.com/inward/record.url?scp=85095120147&partnerID=8YFLogxK
U2 - 10.1007/978-1-0716-0775-6_18
DO - 10.1007/978-1-0716-0775-6_18
M3 - Chapter
C2 - 33128755
AN - SCOPUS:85095120147
SN - 978-1-0716-0774-9
T3 - Methods in Molecular Biology
SP - 251
EP - 284
BT - Protein Downstream Processing
A2 - Labrou, Nikolaos E.
PB - Humana Press
ER -