Abstract
Magnetic particles have been increasingly used as carriers for binding proteins, enzymes, and drugs. Such immobilization procedures for proteins, enzymes, antibodies, and other biologically active compounds have a major impact in different areas of biomedicine and biotechnology. The immobilized biomolecules can be used directly for a bioassay or as affinity ligands to capture or modify target molecules or cells. This chapter details immobilization procedures for proteins and enzymes onto various magnetically responsive carriers such as naked magnetic particles, carboxyl-modified microspheres, and aminomodified microspheres using direct binding procedure in the presence of coupling agents such as carbodiimide. The physical and chemical properties of freshly prepared magnetic particles were determined by magnetic measurements (VSM magnetometer), transmission electron microscopy (TEM), and atomic force microscopy (AFM). The extent of immobilization and enzyme activities were spectrophotometrically measured in order to find the retained activity after immobilization onto magnetic particles. The binding of proteins and enzymes was also confirmed by TEM microscopy.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Reference
Cheng-we W., Jin-gang L., and Wen-chien L. (1998) Protein and enzyme immobilization on non-porous microspheres of polystyrene. Biotechnol. Appl. Biochem. 27, 225–230.
Maa Y. F. and Horvath, C. J. (1988) Rapid analysis of proteins and peptides by reversed-phase chromatography with polymeric micropellicular sorbents. J. Chromatogr. A. 445(1), 71–86.
Rounds M. A. and Regnier F. E. (1988) Synthesis of a non-porous, polystyrenebased strong anion-exchange packing material and its application to fast highperformance liquid chromatography of proteins. J. Chromatogr. A. 443, 73–83.
Wongyai S., Varga J. M., and Bonn G. K. J. (1991) High-performance affinity chromatography of immunoglobulin E on a column of dinitrophenylamino acids covalently bound to a highly cross-linked polymeric micropellicular support. J. Chromatogr. A. 536, 155–164.
Tuncel A., Denizli A., Purvis D., Lowe C. R., and Piskin E. (1993) Cibacron Blue F3G-A-attached monosize poly(vinyl alcohol)-coated polystyrene microspheres for specific albumin adsorption. J. Chromatogr. A. 634, 161–168.
Lee W. C., Lin C. H., Ruaan R. C., and Hsu K. Y. (1995) High-performance affinity chromatography of proteins on non-porous polystyrene beads. J. Chromatogr. A. 704, 307–314.
Smidsrod O., and Skjak-Braek G. (1990) Alginate as immobilization matrix for cells. Trends Biotechnol. 8, 71–78.
Saiyed Z. M., Telang S. D., and Ramchand C. N. (2003) Application of magnetic techniques in the field of drug discovery and biomedicine. BioMagn. Res. Technol. 1, 2.
Ramchand C. N., Pande P., Kopcansky P., and Mehta R. V. (2001) Application of magnetic fluids in medicine and biotechnology. Indian J. Pure Appl. Phys. 39, 683–686.
Mosbach K., and Andersson L. (1977) Magnetic ferrofluids for preparation of magnetic polymers and their application in affinity chromatography. Nature 270, 259–261.
Safarikova M., Roy I., Gupta M. N., and Safarik I. (2003) Magnetic alginate microparticles for purification of a-amylases. J. Biotechnol. 105, 255–260.
Sinclair B. (1998) To bead or not to bead: applications of magnetic bead technology. Scientist 12(13), 17.
Safarik I., and Safarikova M. (2002) Magnetic nanoparticles and biosciences. Mon. Chem. 133, 737–759.
Miltenyi Biotec. Available athttp://www.miltenyibiotec.com.
Polysciences Inc.Available at http://www.polysciences.com..
Abudiab T. and Beitle, R. R. Jr. (1998) Preparation of magnetic immobilized metal affinity separation media and its use in the isolation of proteins. J. Chromatogr. A. 795(2), 211–217.
Sinclair B. (2000) Honing your cloning: new cloning systems give protein expression studies a boost. Scientist 14(16), 29.
QIAGEN Inc.Available athttp://www.qiagen.com/literature/qiagennews/0498/984ninta.pdf.
Safarik I. and Safarikova, M. (2000) Biologically active compounds and xenobiotics: magnetic affinity separations. In: Encyclopedia of Separation Science (Wilson I. D., Adlard T. R., Poole C. F., and Cool M., eds.) AcademicPress, London, pp. 2163–2170.
Mehta R. V., Upadhyay R. V., Charles S. W., and Ramchand C. N. (1997) Direct binding of protein to magnetic particles. Biotechnol. Tech. 11(7), 493–496.
Bacri J., Perzynski R., and Salin D. (1990) Ionic ferrofluids: a crossing of chemistry and physics. J. Magn. Magn. Mater. 85, 27–32.
Koneracka M., Kopcansky P., Timko M., Ramchand C. N., de Sequeira A., and Trevan M. (2002) Direct binding procedure of proteins and enzymes to fine magnetic particles. J. Mol. Catal. B-Enzym. 689, 1–6.
Chantrell R. W. Popplewell J., and Charles S.W. (1978) Measurements particle size distribution parameters in ferrofluids. IEEE Trans. Magn. Mag. 14, 975–977.
Lacava B. M., Azevedo R. B. Silva L. P., et al. (2000) Particle sizing of magnetite-based magnetic fluid using atomic force microscopy: A comparative study with electron micros-copy and birefringence. Appl. Phys. Lett. 77, 1876–1878
Bangs Laboratories Inc.Available ar [http://www.bangslabs.com] and [http://www.bangslabs.com/technotes/205.pdf].
Slavik K. and Smetana R. (1952) Chem. Listy 46, 649.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Koneracká, M. et al. (2006). Immobilization of Enzymes on Magnetic Particles. In: Guisan, J.M. (eds) Immobilization of Enzymes and Cells. Methods in Biotechnology™, vol 22. Humana Press. https://doi.org/10.1007/978-1-59745-053-9_19
Download citation
DOI: https://doi.org/10.1007/978-1-59745-053-9_19
Publisher Name: Humana Press
Print ISBN: 978-1-58829-290-2
Online ISBN: 978-1-59745-053-9
eBook Packages: Springer Protocols