Antibodies specialized programs
Chicken immunization
Using non-mammalian hosts like chicken allows obtaining good immune responses when the antigen is a highly conserved mammalian protein. Moreover, chicken antibodies are present at high quantities in the egg yolk, allowing a non-invasive antibody collection.
Why produce your antibodies in Chicken?
Chicken produce IgY, which migrate from the hen serum to the egg yolk.
Compared to mammalian IgGs, they have one extra Fc domain
IgY’s do not bind to protein A or G, and do not cross-react with anti-mammal secondary antibodies (anti-rabbit IgG, anti-mouse IgG…). This offers a high advantage for double staining experiments, where one can combine one IgG with one IgY, both lacking any cross-reaction to the secondary antibody of the other immunoglobulin type.
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Specificity
Allows targeting conserved mammalian proteins
Specificity
Due to the phylogenetic gap between mammals and birds, chicken may be immunized with proteins highly conserved across mammalian
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Sensitivity
Gives less background
Sensitivity
IgY’s do not bind to the Fc receptors on the surface of mammalian cells, highly reducing background in several applications
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High quantity
Eggs can be collected at high yields
Large quantity
IgY’s may be collected from multiple eggs of a same animal, leading to high batch quantities; Frozen egg yolk and eggs stored at 4°C are stable products. Four eggs contain as much antibody as the serum from one rabbit (200 mg total IgY, 2-10 % being antigen-specific).
Chicken Immunization
Programs overview
Several antigens such as recombinant proteins and peptides may be used, provided by the customer or produced by Eurogentec.
If using peptides, ‘Double X programs’, where a combination of 2 peptides targeting different protein regions are co-injected, will increase the chances of success.
| Anti-antigen | Anti-peptide | Double X | |
|---|---|---|---|
| You provide the antigena (peptide, protein, complex sample) | Eurogentec designsc and synthesises one peptide | Eurogentec designs and synthesises two peptides to increase the chances of success | |
| Peptide synthesis | X | 1 peptide | 2 peptides |
| >70% purity | >70% purity | ||
| 10-16 aa | 10-16 aa | ||
| 15-25 mg | 15-25 mg | ||
| Coupling to carrier | (O) | 5 mg | 5 mg of each peptide |
| Coupled to | KLH | KLH | KLH |
| BSA | BSA | BSA | |
| OVA | OVA | OVA | |
| THY | THY | THY | |
| or MAP carrier | or MAP carrier | or MAP carrier | |
| Number of host | 2 | 2 | 2 |
| ELISA Guaranteeb | X | (O) | (O) |
| Affinity purification | (O) | (O) | (O) |
| QC validation | (O) | (O) | (O) |
| Delivery | Egg yolks | Egg yolks | Egg yolks |
| Purified antibody (O) | Purified antibody (O) | Purified antibody (O) | |
| Remaining peptided | Remaining peptided |
a. See the section "How to provide your antigen"
b. The guarantee only applies for peptides when approved by Eurogentec
c. Please note that peptide design aims at conferring high antigenicity to the peptides, but doesn’t guaranty that they will lead to antibodies functional in any application. To guarantee an optimal functionality of the generated antibodies in your application, we suggest our DoubleX strategy which involves the design and co-injection of two peptides from the same antigen. This co-immunisation more closely mimics the protein than a single peptide strategy and has success rates in the 90 % in Western Blots using this technique.
d. There is no minimum amount of remaining peptide quaranteed after coupling, ELISA test and purification. If a specific quantity of unconjugated peptide is requested, it will be provided as separated aliquots for an additional fee.
X not applicable
(O) optional
