Purification of IgG Using Affinity Chromatography on Antigen-Ligand Columns

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

• 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. 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).