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Purification of IgG Using Affinity Chromatography on Antigen-Ligand Columns

  • Mark Page
  • Robin Thorpe
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Affinity chromatography is a particularly powerful procedure, which can be used to purify IgG, subpopulations of IgG, or the antigen binding fraction of IgG present in serum/ascitic fluid/hybridoma culture supernatant. This technique requires the production of a solid matrix to which a ligand having either affinity for the relevant IgG or vice versa has been bound (1). Examples of ligands useful in this context are:
  1. 1.

    The antigen recognized by the IgG (for isolation of the antigen-specific fraction of the serum/ascitic fluid, and so forth).

     
  2. 2.

    IgG prepared from an anti-immunoglobulin serum, e.g., rabbit antihuman IgG serum or murine antihuman IgG MAb for the purification of human IgG (see Note 1).

     
  3. 3.

    IgG-binding proteins derived from bacteria, e.g., protein A (from Staphylococcus aureus Cowan 1 strain) or proteins G or C (from Streptococcus and see Chapter 130).

     

Keywords

Sodium Citrate Buffer Fume Hood Guanidine Hydrochloride Trisodium Citrate Cyanogen Bromide 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Hermanson, G. T., Mallia, A. K., and Smith, P. K. (1992) Immobilized Affinity Ligand Techniques. Academic, San Diego, CA.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1996

Authors and Affiliations

  • Mark Page
    • 1
  • Robin Thorpe
    • 2
  1. 1.MEDEVA, Vaccine Research Unit, Department of BiochemistryImperial College of Science, Technology, and MedicineLondonUK
  2. 2.National Institute for Biological Standards and ControlPotters BarUK

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