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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Groarke R, Brabazon D (2016) Methacrylate polymer monoliths for separation applications. Materials 9:446
González-González M, González-Valdez J, Mayolo-Deloisa K, Rito-Palomares M (2017) Monolithic chromatography: insights and practical perspectives. J Chem Technol Biotechnol 92:9–13
Pfaunmiller EL, Paulemond ML, Dupper CM, Hage DS (2013) Affinity monolith chromatography: a review of principles and recent analytical applications. Anal Bioanal Chem 405:2133–2145
Švec F, Tennikova TB, Deyl Z (2003) Monolithic materials : preparation, properties and applications. Elsevier, Amsterdam
Satzer P, Sommer R, Paulsson J, Rodler A, Zehetner R, Hofstädter K, Klade C, Jungbauer A (2018) Monolith affinity chromatography for the rapid quantification of a single-chain variable fragment immunotoxin. J Sep Sci 41:3051–3059
Lendero Krajnc N, Podgornik A, Štrancar A, Černigoj U, Nemec B, Vidic U, Vidič J, Gašperšič J (2016) Characterization of methacrylate chromatographic monoliths bearing affinity ligands. J Chromatogr A 1464:72–78
Naldi M, Tramarin A, Bartolini M (2018) Immobilized enzyme-based analytical tools in the -omics era: recent advances. J Pharm Biomed Anal 160:222–237
Han X, Xie Y, Wu Q, Wu S (2019) The effect of monolith properties on the digestion performance of monolith-based immobilized enzyme microreactor. J Chromatogr Sci 57:116–121
Vlakh EG, Tennikova TB (2013) Flow-through immme reactors based on monoliths: I. Preparation of heterogeneous biocatalysts. J Sep Sci 36:110–127
Ralla K, Anton F, Scheper T, Kasper C (2009) Application of conjoint liquid chromatography with monolithic disks for the simultaneous determination of immunoglobulin G and other proteins present in a cell culture medium. J Chromatogr A 1216:2671–2675
Volokitina MV, Bobrov KS, Piens K, Eneyskaya EV, Tennikova TB, Vlakh EG, Kulminskaya AA (2015) Xylan degradation improved by a combination of monolithic columns bearing immobilized recombinant β-xylosidase from Aspergillus awamori X-100 and Grindamyl H121 β-xylanase. Biotechnol J 10:210–221
Milačič R, Zuliani T, Vidmar J, Ščančar J (2016) Monolithic chromatography in speciation analysis of metal-containing biomolecules: a review. J Anal At Spectrom 31:1766–1779
Waterborg JH, Matthews HR (2009) The Lowry method for protein quantitation. In: Walker JM (ed) The protein protocols handbook. Humana Press INC, Totowa, NJ, USA, p 1984
Ostryanina ND, Vlasov GP, Tennikova TB (2002) Multifunctional fractionation of polyclonal antibodies by immunoaffinity high-performance monolithic disk chromatography. J Chromatogr A 949:163–171
Somogyi M (1952) Notes on sugar determination. J Biol Chem 195:19–23
Platonova GA, Vlakh EG, Ivanova ND, Tennikova TB (2009) A flow-through enzymatic bioreactor based on immobilized α-chymotrypsin. Russ J Appl Chem 82(12):2182–2186
Platonova GA, Surzhik MA, Tennikova TB, Vlasov GP, Timkovskii AL (1999) The catalysis of polyriboadenylate synthesis and phosphorolysis by polynucltotide phosphorylase immobilized on a new type of carrier. Russ J Bioorgan Chem 25:166–171
Wang X, Xia D, Han H, Peng K, Zhu P, Crommen J, Wang Q, Jiang Z (2018) Biomimetic small peptide functionalized affinity monoliths for monoclonal antibody purification. Anal Chim Acta 1017:57–65
Vlakh E, Novikov A, Vlasov G, Tennikova T (2004) Solid phase peptide synthesis on epoxy-bearing methacrylate monoliths. J Pept Sci 10:719–730
Meller K, Pomastowski P, Szumski M, Buszewski B (2017) Preparation of an improved hydrophilic monolith to make trypsin-immobilized microreactors. J Chromatogr B 1043:128–137
Ponomareva EA, Kartuzova VE, Vlakh EG, Tennikova TB (2010) Monolithic bioreactors: effect of chymotrypsin immobilization on its biocatalytic properties. J Chromatogr B 878:567–574
Han W, Yamauchi M, Hasegawa U, Noda M, Fukui K, van der Vlies AJ, Uchiyama S, Uyama H (2015) Pepsin immobilization on an aldehyde-modified polymethacrylate monolith and its application for protein analysis. J Biosci Bioeng 119:505–510
Vlakh EG, Tennikova TB (2013) Flow-through immobilized enzyme reactors based on monoliths: I. preparation of heterogeneous biocatalysts. J Sep Sci 36:110–127
Vlakh EG, Platonova GA, Vlasov GP, Kasper C, Tappe A, Kretzmer G, Tennikova TB (2003) In vitro comparison of complementary interactions between synthetic linear/branched oligo/poly-L-lysines and tissue plasminogen activator by means of high-performance monolithic-disk affinity chromatography. J Chromatogr A 992:128–138
Platonova GA, Pankova GA, Il'ina IY, Vlasov GP, Tennikova TB (1999) Quantitative fast fractionation of a pool of polyclonal antibodies by immunoaffinity membrane chromatography. J Chromatogr A 852:129–140
Gupalova TV, Lojkina OV, Palagnuk VG, Totolian AA, Tennikova TB (2002) Quantitative investigation of the affinity properties of different recombinant forms of protein G by means of high-performance monolithic chromatography. J Chromatogr A 949:185–193
Vlakh EG, Volokitina MV, Vinokhodov DO, Tennikova TB (2014) Degradation of polyribonucleotides: biocatalysis and the monitoring of products. Appl Biochem Microbiol 50:600–607
Ponomareva EA, Kartuzova VE, Vlakh EG, Tennikova TB (2010) Monolithic bioreactors: effect of chymotrypsin immobilization on its biocatalytic properties. J Chromatogr B Anal Technol Biomed Life Sci 878:567–574
Vlakh E, Ostryanina N, Jungbauer A, Tennikova T (2004) Use of monolithic sorbents modified by directly synthesized peptides for affinity separation of recombinant tissue plasminogen activator (t-PA). J Biotechnol 107:275–284
Vlakh EG, Tappe A, Kasper C, Tennikova TB (2004) Monolithic peptidyl sorbents for comparison of affinity properties of plasminogen activators. J Chromatogr B Anal Technol Biomed Life Sci 810:15–23
Kalashnikova IV, Ivanova ND, Evseeva TG, Menshikova AY, Vlakh EG, Tennikova TB (2007) Study of dynamic adsorption behavior of large-size protein-bearing particles. J Chromatogr A 1144:40–47
Vlakh EG, Ponomareva EA, Tennikova TB (2014) A multienzyme bioreactor based on a chitinase complex. Appl Biochem Microbiol 50:441–446
Volokitina MV, Nikitina AV, Tennikova TB, Korzhikova-Vlakh EG (2017) Immobilized enzyme reactors based on monoliths: effect of pore size and enzyme loading on biocatalytic process. Electrophoresis 38:2931–2939
Raines RT, Ribonuclease A (1998) Chem Rev 98:1045–1065
Ma W, Tang C, Lai L (2005) Specificity of trypsin and chymotrypsin: loop-motion-controlled dynamic correlation as a determinant. Biophys J 89:1183–1193
By H, Bpn S (1975) Use of IV-benzoyl-L-tyrosine substrate Ester as a protease. J Biol Chem 250:7366–7371
Dahiya N, Tewari R, Hoondal GS (2006) Biotechnological aspects of chitinolytic enzymes: a review. Appl Microbiol Biotechnol 71:773–782
Sousa S, Ramos A, Evtuguin DV, Gamelas JAF (2016) Xylan and xylan derivatives—their performance in bio-based films and effect of glycerol addition. Ind Crop Prod 94:682–689
Ponomareva EA, Volokitina MV, Vinokhodov DO, Vlakh EG, Tennikova TB (2013) Biocatalytic reactors based on ribonuclease a immobilized on macroporous monolithic supports. Anal Bioanal Chem 405:2195–2206
Martinović T, Andjelković U, Klobučar M, Černigoj U, Vidič J, Lučić M, Pavelić K, Josić D (2017) Affinity chromatography on monolithic supports for simultaneous and high-throughput isolation of immunoglobulins from human serum. Electrophoresis 38:2909–2913
Vlakh EG, Tennikova TB (2013) Flow-through immobilized enzyme reactors based on monoliths: II. Kinetics study and application. J Sep Sci 36:110–127
Mao Y, Černigoj U, Zalokar V, Štrancar A, Kulozik U (2017) Production of β-Lactoglobulin hydrolysates by monolith based immobilized trypsin reactors. Electrophoresis 38:2947–2956
Masini JC, Svec F (2017) Porous monoliths for on-line sample preparation: a review. Anal Chim Acta 964:24–44
Kent UM (1999) Purification of antibodies using ammonium sulfate fractionation or gel filtration. In: Javois LC (ed) Methods in molecular biology: immunocytochemical methods and protocols. Humana Press Inc, Totowa, NJ, pp 11–18
Crowther JR (2009) The ELISA guidebook. In: Methods in molecular biology. Humana Press, Totowa, NJ, p 516
Strancar A, Barut M, Podgornik A, Koselj P, Josic D, Buchacher A (1998) Polymer based supports for fast separation of biomolecules. LC-GC Int 11:660–670
Kasper C, Meringova L, Freitag R, Tennikova T (1998) Fast isolation of protein receptors from streptococci G by means of macroporous affinity disks. J Chromatogr A 798:65–72
Kurien BT, Scofield RH (2012) Protein Electrophoresis. In: Methods in molecular biology. Humana Press, Totowa, NJ, p 869
Gravanis I, Tsirka SE (2008) Tissue-type plasminogen activator as a therapeutic target in stroke. Expert Opin Ther Targets 12:159–170
Thiebaut AM, Gauberti M, Ali C, Martinez De Lizarrondo S, Vivien D, Yepes M, Roussel BD (2018) The role of plasminogen activators in stroke treatment: fibrinolysis and beyond. Lancet Neurol 17:1121–1132
Khan AL, Heys SD, Eremin O (1995) Synthetic polyribonucleotides: current role and potential use in oncological practice. Eur J Surg Oncol 21:224–227
Naldi M, Černigoj U, Štrancar A, Bartolini M (2017) Towards automation in protein digestion: development of a monolithic trypsin immobilized reactor for highly efficient on-line digestion and analysis. Talanta 167:143–157
Volokitina MV, Vlakh EG, Platonova GA, Vinokhodov DO, Tennikova TB (2013) Polymer monoliths as efficient solid phases for enzymatic polynucleotide degradation followed by fast HPLC analysis. J Sep Sci 36:2793–2805
Vlakh EG, Maksimova EF, Tennikova TB (2013) Monolithic polymeric sorbents for high-performance chromatography of synthetic polymers. Polym Sci Ser B 55:55–62
Moussaoui M, Nogués MV, Guasch A, Barman T, Travers F, Cuchillo C (1998) The subsites structure of bovine pancreatic Ribonuclease a accounts for the abnormal kinetic behavior with Cytidine 2′,3′-cyclic phosphate. J Biol Chem 273:25565–25572
Bartolini M, Greig NH, Yu Q, Andrisano V (2009) Immobilized butyrylcholinesterase in the characterization of new inhibitors that could ease Alzheimer’s disease. J Chromatogr A 1216:2730–2738
Acknowledgments
The authors are very grateful to BIA Separations for long-term fruitful cooperation.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
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
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0775-6_18
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0774-9
Online ISBN: 978-1-0716-0775-6
eBook Packages: Springer Protocols