Immunoafflnity Chromatography

  • George W. Jack
Part of the Methods in Molecular Biology™ book series (MIMB, volume 11)


Immunoaffinity chromatography (IAC) harnesses the specificity and avidity of the interaction between an antigen and its antibody to purify the antigen. The technique may be used with either polyclonal or monoclonal antibodies (MAb) (1), but MAbs are preferable for a number of reasons. Polyclonal sera are never specific for a single antibody but reflect the variety of immunological challenges sustained by an animal since birth. Furthermore, the best polyclonal sera are generated using highly purified antigen, so if a purification method already exists, why raise sera to develop a second purification method? Perhaps the main disadvantage of polyclonal sera lies in the wide range of avidity of antibodies they contain; although this ensures that such antibodies will bind their antigen, the dissociation of the antigen-antibody complex may prove impossible using conditions that retain the activity of both.


Binding Buffer Fast Protein Liquid Chromatography Cyanate Ester Immunoaffinity Chromatography Coupling Buffer 
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  1. 1.
    Kohler G. and Milstein C. (1975) Continuous culture of fused cells secreting antibody of predefined specificity. Nature 250, 495–497.CrossRefGoogle Scholar
  2. 2.
    Cuatrecasas P. (1970) Protein purification by affinity chromatography: Derivatisation of agarose and polyacrylamide beads. J. Biol. Chm. 245, 3059–3065.Google Scholar
  3. 3.
    Chase H. A. (1984) Affinity separations utilising imunobilised monoclonal antibodies-a new tool for the biochemical engineer. chem. Eng. Sci. 39, 1099–1125.CrossRefGoogle Scholar
  4. 4.
    Jack G. W., Blazek R., James K., Boyd J. E., and Micklem L. R. (1987) The automated production by immunoaffinity chromatography of the human pitutary glycoprotein hormones thyrotropin, folhtropin and lutropin. J. Chem. Tech. Biotechnol. 38, 45–58.Google Scholar
  5. 5.
    Bazin H. and Malache J.-M.(1986) Rat (and mouse) monoclonal antibodies. V.A simple automated technique of antigen purification by immunoaffinity chromatography. J. Immunol. Methods. 88, 19–24.PubMedCrossRefGoogle Scholar
  6. 6.
    Eveleigh J. W. (1982) Practical considerations in the use of immunosorbents and associated instrumentation. Anal. Chem. Symp. Ser. 9, 293–303.Google Scholar
  7. 7.
    Jack G. W. and Blazek R. (1987) The purification of human thyroid-stimulating hormone by immunoaffinity chromatography. J, Chem. Tech. Biotechnol. 39, 1–10.Google Scholar
  8. 8.
    Staehelin T., Hobbs D. S., Kung H-F., Lai C.-Y., and Pestka S. (1981) Purification and characterisation of recombinant human leukocyte interferon (IFL + A) with monoclonal antibodies. J Biol. Chem. 256, 9750–9754.PubMedGoogle Scholar
  9. 9.
    Jack G. W. and Gilbert H. J. (1984) The purification by immunoaffinity chromatography of bacterial methionyl-(human somatatropin). Biochem. Soc. Trans. 12, 246, 247.Google Scholar
  10. 10.
    Hornsey V. S., Griffin B. D., Pepper D. S., Micklem L. R., and Prowse C. V. (1987) Immunoaffinity Purification of Fractor VIII Complex. Thromb. Haemostas. 57, 102–105.Google Scholar
  11. 11.
    Kowal R. and Parsons R.C. (1980) Stabilisation of proteins immobilised on Sepharose from leakage by glutaxaldehyde crosslinking. Anal. Biochem. 102, 72–76.PubMedCrossRefGoogle Scholar
  12. 12.
    Jack G. W. (1990) The use of immunoaffinity chromatography in the preparation of therapeutic products, in Focus on Laboratory Methods in Immunology, Vol. 2 (Zola H., ed.) CRC, Boca Raton, FL, pp. 127–146.Google Scholar
  13. 13.
    Goding J. W. (1978) Use of staphylococcal Protein A as an immunological reagent. J. Immunol. Method. 20, 241–253.CrossRefGoogle Scholar
  14. 14.
    Bjorck L. and Kronvall G. (1984) Purification and some properues of streptococcal Protein G, a novel IgG-binding reagent. J. Immunol. 133, 969–974.PubMedGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1992

Authors and Affiliations

  • George W. Jack
    • 1
  1. 1.Centre for Applied Microbiology and ResearchPorton Down, SalisburyUK

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