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Purification of Antibodies

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Allergy Methods and Protocols

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 138))

Abstract

Immunoglobulins are a heterogeneous group of proteins. It naturally follows that the strategies for purifying them are diverse and numerous. A good knowledge of their respective physiochemical properties will obviously make the task easier. The choice between using polyclonal and/or monoclonal antibodies will govern the basic approach. Each approach will present its own advantages/disadvantages including cost, ability to produce a high yield, quality, and a need for standardization. The context in which the antibodies will be used is another important aspect to consider. When the demand is for establishing “ultrasensitive” assays, optimal purity and specificity is obviously required.

This chapter will focus on the purification of mammalian IgG from polyclonal (i.e., rabbit) and monoclonal (i.e., mouse sources). IgG is the principal immunoglobulin constituent of mammalian sera. In older animals, it may well represent >80% of the total Ig concentration, because of its higher rate of synthesis and longer half-life.

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References

  1. Cohn, E. J. and Edsall, J. T. (eds.) (1943) Proteins, Amino Acids and Peptides. Reinhold, New York.

    Google Scholar 

  2. Czok, R. and Bücher, T. (1960) Crystallized enzymes from the myogen of rabbit skeletal muscle. Adv. Protein Chem. 15, 315–415.

    Article  CAS  PubMed  Google Scholar 

  3. Dixon, M. and Webb, E. C. (1961) Enzyme fractionation by salting out; a theoretical note. Adv. Protein Chem. 16, 197–219.

    Article  CAS  PubMed  Google Scholar 

  4. Hornsey, V. S., Prowse, C. V., and Pepper, D. S. (1986) Reductive amination for solid-phase coupling of protein. A practical alternative to cyanogen bromide. J. Immunol. Methods 93, 83–88.

    Article  CAS  PubMed  Google Scholar 

  5. Domen, P., Nevens, J., Mallia, K., Hermanson, G., and Klenk, D. (1990) Site directed immobilization of proteins. J. Chromatogr. 510, 293–302.

    Article  CAS  PubMed  Google Scholar 

  6. Hermanson, G. T., Mallia, K. A., and Smith, P. K. (1992) Immobilized Affinity Ligand Techniques. Academic Press, Inc. San Diego, CA.

    Google Scholar 

  7. Steinbuch, M. and Audran, R. (1969) The isolation of IgG from mammalian sera with the aid of caprylic acid. Arch. Biochem. Biophys. 134, 279–284.

    Article  CAS  PubMed  Google Scholar 

  8. Affinity Chromatography Handbook; Principles and Methods. GE Healthcare, Product Code 18-1022-29, pp. 609.

    Google Scholar 

  9. O’Shanessy, D. J., Dobersen, M. J., and Quarles, R. H. (1984) A novel procedure for labeling immunoglobulins by conjugation to oligosaccharide moieties. Immunol. Lett. 8, 273–277.

    Article  Google Scholar 

  10. O’Shanessy, D. J. and Quarles, R. H. (1987) Labeling of the oligosaccharide moieties of immunoglobulins. J. Immunol. Methods 99, 153–161.

    Article  Google Scholar 

  11. Hoffman, W. L. and O’Shanessy, D. J. (1988) Site-specific immobilization of antibodies by their oligosaccharide moieties to new hydrazide derivatized supports. J. Immunol. Methods 112, 113–120.

    Article  CAS  PubMed  Google Scholar 

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© 2008 Humana Press Inc., Totowa, NJ

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Larsson, P.H. (2008). Purification of Antibodies. In: Jones, M.G., Lympany, P. (eds) Allergy Methods and Protocols. Methods in Molecular Medicine, vol 138. Humana Press. https://doi.org/10.1007/978-1-59745-366-0_16

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  • DOI: https://doi.org/10.1007/978-1-59745-366-0_16

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-896-7

  • Online ISBN: 978-1-59745-366-0

  • eBook Packages: Springer Protocols

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