- Published: Wednesday, 07 May 2014 18:38
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IgY, purified from chicken egg yolk and IgG present in mammalian sera contain antibody specificities that are present in the animal's circulation at the time the eggs were formed or at the time the animal was bled. The IgY that you receive from our standard protocol is approximately 90% pure IgY, containing all the IgY specificities that are present in the hen's circulation and transferred to her egg.
The specific activity (percentage of immunoglobulins that is directed against your antigen) of this IgY fraction (or animal serum) depends on the immunogenicity of your antigen and the genetic make-up of the immunized animal. It is reasonable to expect between 2 and 10% specific activity of IgY/IgG from an animal immunized with a protein greater than 50 kilodaltons. Expect less than 1% specific activity to a peptide from an animal immunized with peptide KLH (there will be a much higher specific activity directed to the carrier KLH portion of the immunogen). For many applications, this non-affinity purified antibodiy fraction will work just fine and it is unnecessary to further purify your IgY/IgG. In fact, further purification may result in the loss of your highest avidity antibodies.
However, sometimes, it is necessary to remove the antibodies that are not antigen specific and this is done by making an antigen column to which the whole IgY/IgG fraction is applied. The IgY/IgG you receive is the antigen-specific fraction that is eluted from the column. To make the affinity column, we will need 5 mg protein antigen (in addition to that supplied for immunizations) at a concentration greater than 3 mg/ml.
Gallus Immunotech Inc. specializes in the development and purification of IgY antibodies, primarily in chickens. However, if you have animal sera from which you would like affinity purified antibodies, we would be happy to do this for you.
If you want affinity-purified IgY from 2 hens immunized with a peptide that we synthesize and conjugate to KLH, please see our Affinity Purifed anti-Peptide Combo for special pricing.
Our custom affinity purification service includes:
- making an affinity column with your antigen or peptide
- antibody affinity purification of antigen- or peptide-specific IgY (or IgG) from a 0.5 gram purified chicken IgY antibody fraction or 25 ml animal serum sample: