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IgY Polyclonal Antibodies: Immunization Protocol

The following is an example of an immunization protocol that has been used for the successful generation of IgY polyclonal antibodies:

  1. Collect a couple of eggs before immunization begins. IgY purified from these eggs will serve as your control IgY.
  2. On day 0, inject between 0.02 and 0.5 mg antigen (with Freund’s complete adjuvant) subcutaneously and/or intramuscularly into the breast tissue of the hen at multiple sites. The total volume of antigen /adjuvant should be about 1 ml with the adjuvant making up between half and two-thirds the volume. Use a comparable amount of antigen that you would use to immunize a rabbit.
  3. Repeat immunizations on day 10, 20 and 30 using incomplete Freund’s adjuvant and about half the amount of antigen. Specific antibody should be detected by day 30 in the eggs. For prolonged antibody production, hens should receive boosts every couple of months.
  4. Individual hens will react differently to immunization with a particular antigen. Antibody titres will vary as will the time period in which the titres are stable.

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