Method for immunizing an avian species
The present invention relates to a method for generating a plurality of different IgY antibodies, by immunizing each of a plurality of avian organisms of the same or different species with a composition comprising a plurality of different antigens, thereby generating a plurality of different IgY antibodies.
1. A method for generating a library of randomly generated IgY antibodies for targeting multiple ligands, said library comprising a plurality of antibodies, said method comprising the steps of:
a) generating a peptide library by random peptide synthesis, wherein the resulting peptides are randomized at each amino acid residue and at least the 20 naturally occurring amino acids are represented at each amino acid position of peptides in the library, and wherein the length of the resulting peptides is between 5 and 15 amino acid residues,
b) immunizing each of a plurality of avian organisms of the same or different species with a composition comprising a plurality of different peptides of the library produced in step a),
c) isolating antibodies from the yolk of eggs laid by the plurality of immunized avian organisms of step b), and
d) pooling said isolated antibodies.
2. A method for generating IgY antibodies for targeting to a ligand, in which method the ligand is not used for immunization, said method comprising the steps of:
a) generating a peptide library by random peptide synthesis, wherein the resulting peptides are randomized at each amino acid residue and at least the 20 naturally occurring amino acids are represented at each amino acid position of peptides in the library, and wherein the length of the resulting peptides is between 5 and 15 amino acid residues,
b) immunizing each of a plurality of avian organisms of the same or different species with a composition comprising a plurality of different peptides of the library produced in step a),
c) isolating antibodies from the yolk of eggs laid by the plurality of immunized avian organisms of step b),
d) pooling said isolated antibodies,
e) targeting said pool of antibodies with the ligand, and
f) isolating a library of antibodies having affinity for said ligand.
3. The method according to claim 1 , wherein the ligands are not used for immunization.
4. The method according to any one of claims 1 - 3 , wherein in step b) each avian organism is immunized with at least about 100 different peptides.
5. The method according to any one of claims 1 - 3 , wherein in step b) a total of from 10 to 100000 avian organisms are immunized.
6. The method according to claim 5 , wherein between 600 and 7000 avian organisms are immunized.
7. The method according to any one of claims 1 - 3 , wherein the avian organism is a bird of the gallinacean type.
8. A method for isolating one or more IgY antibodies with affinity for a ligand without prior immunization with said ligand, said method comprising the steps of:
a) generating a library of IgY antibodies by the method of claim 1 to yield a pool of IgY antibodies,
b) targeting said pool of antibodies with the ligand, and
c) isolating one or more antibodies having affinity for said ligand.
9. The method according to claim 8 , wherein each avian species is immunized with at least about 100 different peptides.
10. The method according to claim 8 , wherein a total of from 10 to 100000 avian organisms are immunized.
11. The method according to claim 10 , wherein a total of between 600 and 7000 avian organisms are immunized.
12. The method according to claim 8 , wherein the avian organism is a bird of the gallinacean type.