Design and fabrication of superparamaganitic hybrid microspheres for protein immobilization

Abstract

Superparamagnetic poly(styrene)-co-poly(2-acrylanmido-2-methyl propanesulfonic acid) (PSt-co-PAMPS) and poly(methylmethacrylate)-co-poly(glycidyl methacrylate) (PMMA-co-PGMA) microspheres with mean size of 170 nm were prepared by emulsion polymerization in the presence of oleic acid-coated Fe₃O₄ nanoparticles. The structures, morphologies, diameter and diameter distribution of the as-prepared microspheres were identified by Fourier transform infrared spectroscopy (FT-IR) and transmission electron microscopy (TEM). The saturation magnetizations of PSt-co-PAMPS and PMMA-co-PGMA microspheres are 21.94 and 25.07 emu/g, respectively. The as-synthesized magnetic microspheres were used for immobilization of Bovine serum albumin (BSA) by physical interaction and covalent interaction respectively. The equilibrium amount of BSA immobilized onto PMMA-co-PGMA microspheres was 86.48 mg/g microspheres in 90 min, while on PSt-co-PAMPS microspheres was 59.62 mg/g microspheres in 120 min.

Keywords

Fe₃O₄ nanoparticles / emulsion polymerization / magnetic composite microspheres / protein immobilization

References

[1] Huang X. L., Zhuang J., Chen D., . General Strategy for Designing Functionalized Magnetic Microspheres for Different Bioapplications[J]. Langmuir., 2009, 25(19): 11 657-11 663.

[2] Qiu G. M., Xu Y. Y., Zhu B. K., . Novel, Fluorescent, Magnetic, Polysaccharide-Based Microsphere for Orientation, Tracing, and Anticoagulation: Preparation and Characterization[J]. Biomacromolecules, 2005, 6(2): 1 041-1 047.

[3] Zhou S. B., Sun J., Sun L., . Preparation and Characterization of Interferon-loaded Magnetic Biodegradable Microspheres[J]. J. Biomed. Mater. Res.-Appl. Biom., 2008, 87B(1): 189-196.

[4] Xie X., Zhang X., Zhang H., . Preparation and Application of Surface-coated Superparamagnetic Nanobeads in the Isolation of Genomic DNA[J]. J. Magn. Magn. Mater., 2004, 277: 16-23.

[5] Fang H., Huang J., Ding L. Y., . Preparation of Magnetic Chitosan Nanoparticles and Immobilization of Laccase[J]. Journal of Wuhan University of Technology-Mater. Sci. Ed., 2009, 24(1): 42-47.

[6] Yang Y., Yong X. B., Li Y. F., . Characterization of Candida Rugosa Lipase Immobilized onto Magnetic Microspheres with Hydrophilicity[J]. Process Biochem., 2008, 43: 1 179-1 185.

[7] Bahar T., Tuncel A. Immobilization of a-chymotrypsin onto Newly Produced Poly(hydroxypropyl methacrylate -comethacrylic acid) Hydrogel Beads[J]. React. Funct. Polym., 2000, 44: 7-78.

[8] Guo N., Wu D. C., Pan X. H., . Magnetic Polymer Microspheres with Azidocarbonyl Groups: Synthesis, Characterization and Application in Protein Immobilization[J]. J. Appl. Polym. Sci., 2009, 112: 2 383-2 390.

[9] Hayens C. A., Norde W. Structures and Stabilities of Absored Protein[J]. J. Colloid. and Interface Sci., 1995, 169: 313-328.

[10] Fang S. J., Kawaguchi H. Colloidal Properties and Protein Adsorption of Amphoteric Microspheres[J]. Colloids and Surfaces A: Physicochem. Eng. Aspects, 2002, 211: 79-84.

[11] Arshady R. Microspheres for Biomedical Applications: Preparation of Reactive and Labelled Microspheres[J]. Biomaterials, 1993, 14(1): 5-15.

[12] Gong X, Peng S L, Wen W, . Design and Fabrication of Magnetically Functionalized Core/Shell Microspheres for Smart Drug Delivery[J]. Adv. Funcl. Mater., 2009, 19: 292-297.

[13] Ding X. B., Sun Z. H., Wan G. X. Interaction between Thermosensitive Magnetic Polymer Microspheres and Proteins[J]. Acta Polymerica Sinica (in Chinese), 2000, 1: 9-13.

[14] Han D. K., Park K. D., Ryu G. H., . Plasma Protein Adsorption to Sulfonated Poly(ethy1ene oxidebgrafted polyurethane surface[J]. J. Biomed. Mater. Res., 1996, 30: 23-30.

[15] Ma Z. Y., Guan Y. P., . Covalent Immobilization of Albumin on Micron-sized Magnetic Poly(methyl methacrylatedivinylbenzeneglycidyl methacrylate) Microspheres Prepared by Modified Suspension Polymerization[J]. Polym. Advd. Technol., 2005, 16: 554-558.

[16] Yoon J. Y., Kim J. H., Kim W. S. The Relationship of Interaction Forces in the Protein Adsorption onto Polymeric Microspheres[J]. Colloids and Surfaces A, 1999, 153: 413-419.

[17] Kawaguchi H., Amagasa H., Hagiya T., . Interaction between Proteins and Latex Particles Having Different Surface Structures[J]. Colloids and Surfaces, 1985, 13: 295-311.

[18] Krizova J., Spanova A., Ritticha B., . Magnetic Hydrophilic Methacrylate-based Polymer Microspheres for Genomic DNA Isolation[J]. J Chromatography A, 2005, 1064: 247-253.

[19] Li R. X., Liu S. M., Zhao J. Q., . Application of Polymerizable Surfactant in the Preparation of Polystyrene/Nano-Fe3O4 Composite[J]. Journal of Wuhan University of Technology-Mater. Sci. Ed., 2010, 25(2): 184-187.

[20] Yang C. L., Liu H. Z., Guan Y. P., . Preparation of Magnetic Poly(methylmethacrylate-divinylbenzene-glycidylmethacrylate) Microspheres by Spraying Suspension Polymerization and Their Use for Protein Adsorption[J]. J. Magn. Magn. Mater., 2005, 293: 187-192.

[21] Mackova H., Kralova D., Horak D. Magnetic Poly(Nisopropylacrylamide) Microspheres by Dispersion and Inverse Emulsion Polymerization[J]. J. Polym. Sci.-A: Polym. Chem., 2007, 45: 5 884-5 898.

[22] Zhang J., Zhang S., Wang Y. P., . Composite Magnetic Microspheres: Preparation and Characterization[J]. J. Magn. Magn. Mater., 2007, 309: 197-201.

[23] Park S. W., Kim Y. I., Chung K. H., . Covalent Immobilization of GL-7-ACA Acylase on Silica Gel Through Silanization[J]. React. Funct. Polym., 2002, 51: 79-92.

[24] Sun Y., Wang B., Hui C., . Synthesis and Characterization of Magnetic Polymer Microspheres[J]. J. Macromol. Sci. Part B: Physics, 2006, 45: 653-658.

[25] Gong T., Dong Y., Hua J. H., . Preparation of Monodispersed Hybrid Nanospheres with High Magnetite Content from Uniform Fe₃O₄ Clusters[J]. Colloids and Surfaces A: Physicochem. Eng. Aspects, 2009, 339: 232-239.