Polyethylene Glycol (PEG) and /or Ammonium Sulphate Precipitation

• Protein A Affinity Chromatography

• Protein G Affinity Chromatography

• Protein A/G Affinity Chromatography

• Ig Affinity

• Antigen Affinity

This method is useful for concentration and partial purification of antibodies from most sources and all species including purification of IgY’s from chicken egg yolks. Although, on its own the yields of antibodies are impure, it is the method of choice when combined with ion exchange for large volumes of antisera and or followed by Antigen Affinity purification to enrich for specific antibodies. It is not recommended for purification of tissue culture supernatant.

This process binds the Fc region of major subclasses of mouse and rat IgG to varying degrees. The results with Protein A are usually high purity and high yields. It is useful for large-scale purifications of tissue culture supernatants.

Protein G can be used for purifying antibody from most sources. Like Protein A, the results are usually of high purity and high yield. It is applicable to a wider range of immunoglobulins than Protein A, but elution procedures may be harsher.

This method can be used to isolate monoclonal antibodies from supernatants which contain high levels of serum Ig. It is the choice for secondary antibody production.

This is the method of choice when specific antibody is required from polyclonal antisera. For best results this method requires pure antigen for the preparation of the affinity columns.