Skip to main content
Log in

Development and application of thermo-sensitive magnetic immunomicrospheres for antibody purification

  • Applied Microbial and Cell Physiology
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Ultrafine magnetite particles were prepared by a co-precipitation method. The poly-(styrene/N-isopropylacrylamide/methacrylic acid) latex particles containing ultrafine magnetite [magnetic P(St/NIPAM/MAA)] were prepared by two-step emulsifier-free emulsion polymerization. The minimum NaCl concentration for flocculation of these magnetic latex particles (critical flocculation concentration, CFC) decreased with increasing temperature. These temperature dependence of CFC, namely its thermo-sensitivity, originated from NIPAM. At a certain NaCl concentration, some of the magnetic latex particles showed reversible transition between flocculation and dispersion by controlling the temperature, and the thermo-flocculated magnetic latex particles were separated quickly in a magnetic field. Bovine serum albumin (BSA) was covalently immobilized onto the magnetic P(St/NIPAM/MAA) latex particles with high efficiency by the carbodiimide method. These thermo-sensitive magnetic immunomicrospheres were effective for the immunoaffinity purification of anti-BSA antibodies from antiserum.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Dunnill P, Lilly MD (1974) Purification of enzymes using magnetic bioaffinity materials. Biotechnol Bioeng 16:987–990

    Google Scholar 

  • Fair BD, Jamieson AM (1980) Studies of protein adsorption on polystyrene latex surfaces. J Colloid Interface Sci 77:525–534

    Google Scholar 

  • Heskins M, Guillet JE (1968) Solution properties of poly(N-isopropyl acrylamide). J Macromol Sci Chem A2:1441–1455

    Google Scholar 

  • Hirschbein BL, Whitesides GM (1982) Affinity separation of enzymes from mixtures containing suspended solids. Comparisons of magnetic and nonmagnetic techniques. Appl Biochem Biotechnol 7:157–176

    Google Scholar 

  • Hoshino F, Fujimoto T, Kawaguchi H, Ohtsuka Y (1987) N-Substituted acrylamide-styrene copolymer lactices II. Polymerization behavior and thermosensitive stability of lactices. Polym J 19:241–247

    Google Scholar 

  • Kondo A, Higashitani K (1992) Adsorption of model proteins with wide variation in molecular properties on colloidal particles. J Colloid Interface Sci 150:344–351

    Google Scholar 

  • Kondo A, Yamasaki R, Higashitani K (1992) Affinity purification of antibodies using immunomicrospheres. J Ferment Bioeng 74:226–229

    Google Scholar 

  • Kondo A, Kaneko T, Higashitani K (1993) Purification of fusion proteins using affinity microspheres in aqueous two-phase systems. Appl Microbiol Biotechnol 40:365–369

    Google Scholar 

  • Lea T, Vartdal F, Nustad K, Funderud S, Berge A, Ellingsen T, Schmid R, Stenstad P, Ugelstad J (1988) Monosized, magnetic polymer particles: their use in separation of cells and subcellular components, and in the study of lymphocyte function in vitro. J Mol Recog 1:9–18

    Google Scholar 

  • Molday RS, Mackenzie D (1982) Immunospecific ferromagnetic iron-dextran reagents for the labeling and magnetic separation of cells. J Immunol Methods 52:353–367

    Google Scholar 

  • Mosbach K, Andersson L (1977) Magnetic ferrofluids for preparation of magnetic polymers and their application in affinity chromatography. Nature 270:259–261

    Google Scholar 

  • Norde W, Lyklema J (1978) The adsorption of human plasma albumin and bovine pancreas ribonuclease at negatively charged polystyrene surfaces. I. Adsorption isotherms. Effects of charged, ionic strength, and temperature. J Colloid Interface Sci 66:257–265

    Google Scholar 

  • Platsoucas CD (1987) Biomedical applications of polymer particles with emphasis on cell separation. In: El-Asser MS, Fitch RM (eds) Future directions in polymer colloids. Marutinus Nijhoff, Dordrecht, p. 321

    Google Scholar 

  • Sada E, Katoh S, Kondo A, Kiyokawa A (1986) Effects of coupling method and ligand concentration on adsorption equilibrium in immuno-affinity chromatography. J Chem Eng Jpn 19:502–506

    Google Scholar 

  • Setchell CH (1985) Magnetic separations in biotechnology — a review. J Chem Technol Biotechnol 35B:175–182

    Google Scholar 

  • Shimoiizaka J (1979) Japanese patent no. 54-40069

  • Shinkai M, Honda H, Kobayashi T (1991) Preparation of fine magnetic particles and application for enzyme immobilization. Biocatalysis 5:61–69

    Google Scholar 

  • Suzawa T, Shirahama H, Fujimoto T (1982) Adsorption of bovine serum albumin onto homo- and copolymer latices. J Colloid Interface Sci 86:144–150

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Correspondence to: A. Kondo

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kondo, A., Kamura, H. & Higashitani, K. Development and application of thermo-sensitive magnetic immunomicrospheres for antibody purification. Appl Microbiol Biotechnol 41, 99–105 (1994). https://doi.org/10.1007/BF00166089

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00166089

Keywords

Navigation